Antagonists of IL-6 to prevent or treat thrombosis

ABSTRACT

The present invention is directed to therapeutic methods using IL-6 antagonists such as antibodies and fragments thereof having binding specificity for IL-6 to prevent or treat thrombosis in diseases associated with abnormal blood coagulation or fibrinolysis. In preferred embodiments these patients will comprise those exhibiting elevated D-dimer or other cogulation cascade related proteins and optionally will further exhibit elevated C reactive protein prior to treatment. The subject therapies also may include the administration of other actives such as chemotherapeutics, anti-coagulants, statins, et al.

This application is a divisional of Ser. No. 14/679,767, filed Apr. 6,2015, now U.S. Pat. No. 9,926,370, which is a divisional of U.S.application Ser. No. 13/548,994, filed Jul. 13, 2012, now. U.S. Pat. No.8,999,330, which is a continuation of U.S. application Ser. No.12/323,194, filed Nov. 25, 2008, now U.S. Pat. No. 8,252,286, which is acontinuation-in-part of U.S. application Ser. No. 12/153,612, filed May21, 2008, now U.S. Pat. No. 7,935,340, which claims the benefit of U.S.Provisional Appl. No. 60/924,550, filed May 21, 2007, the disclosures ofeach of all the above listed applications are herein incorporated byreference in their entirety.

The sequence listing in the file named “1156256o001608.txt” having asize of 249,995 bytes that was created Apr. 1, 2020 is herebyincorporated by reference in its entirety.

BACKGROUND OF THE INVENTION Field of the Invention

This invention is an extension of Applicants' prior invention disclosedin the above-referenced patent applications relating to novel anti-IL-6antibodies and novel therapies and therapeutic protocols using anti-IL-6antibodies, preferably those described herein. In particular, thisinvention pertains to methods of preventing or treating thrombosis in apatient in need thereof, comprising administering to the patient an IL-6antagonist, whereby the patient's coagulation profile is improved.

In another embodiment the invention provides novel combination therapieswherein thrombosis is prevented or treated by the administration of atleast one IL-6 antagonist and at least one other therapeutic compounde.g., a statin compound including but not limited to pravastatin,lovastatin, simvastatin, fluvastatin, atorvastatin, simvastatin,nystatin, pentastatin, and cerivastatin.

In another embodiment, this invention relates to methods of preventingor treating thrombosis in a patient in a state of hypercoagulation,comprising administering to the patient an anti-IL-6 antibody orantibody fragment, whereby the patient's coagulation profile is improvedor restored to a normal condition, and monitoring the patient to assesscoagulation profile, wherein the anti-IL-6 antibody or antibody fragmentspecifically binds to the same linear or conformational epitope(s)and/or competes for binding to the same linear or conformationalepitope(s) on an intact human IL-6 polypeptide or antibody fragmentthereof as an anti-IL-6 antibody selected from the group consisting ofAb1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13,Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25,Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, and Ab36.

This invention further pertains to novel methods of preventing ortreating thrombosis in a patient in need thereof using anti-IL-6antibodies, preferably humanized antibodies possessing an eliminationhalf-life which is at least about 25 days.

Description of Related Art

Thrombosis is a significant cause of mortality in cancer patients. Bick,N Engl J Med 349:109-111 (2003). For example, serious, life-threateningthrombotic events occur in approximately 6% of lung cancer patients.Alguire et al., J Clin Oncol 2004 Vol (July 15th Supplement) No. 14S:8082. Cancer patients often exhibit hypercoagulation, in which thecoagulation system has an increased clotting tendency. Rickles andEdwards, Blood 62:14-31 (1983). Markers of hypercoagulation correlatewith poor patient outcome for at least some cancers. Bick, Semin ThrombHemostat 18:353-372 (1992); Buccheri et al., Cancer 97:3044-3052 (2003);Wojtukiewicz, Blood Coagul Fibrinolysis 3:429-437 (1992). Causes ofhypercoagulation include the cancer itself and the cancer treatments(e.g., chemotherapy). Hypercoagulation results in an increased risk ofthrombitic events, which can be further exacerbated when patients becomebed-ridden. When not contraindicated, anticoagulant therapy hasconferred survival benefit in some cancers. Lebeau et al., Cancer74:38-45 (1994); Chahinian et al., J Clin Oncol 7:993-1002 (1989).However, therapeutic options are often limited because many cancerpatients are at an elevated risk of major bleeding, precludingadministration of anticoagulants that could otherwise be givenprophylactically to reduce the risk of thrombosis. In summary, theavailable methods for prevention of thrombosis in cancer patients areunsatisfactory, and thus there is a need for new therapies. Suchtherapies would enhance cancer patient survival and promote betterquality of life.

Thrombosis can also be a significant cause of adverse events andmortality in other patient groups, including those with chronic illnessor chronic inflammation, surgical patients, bed-ridden individuals, andorthopedic patients. When they are not otherwise contraindicated,preventative methods include calf compression and anticoagulants (e.g.low molecular weight heparin). These preventative methods can reduce—butnot eliminate—the risk of thrombosis. Because these preventative methodsare not always effective and are contraindicated for some patients, andbecause anticoagulants can cause potentially lethal side-effects such asmajor bleeding, there is a need for alternative methods to preventthrombosis in these patients. Such methods should improve patientoutcomes.

Interleukin-6 (hereinafter “IL-6”) (also known as interferon-β₂; B-celldifferentiation factor; B-cell stimulatory factor-2; hepatocytestimulatory factor; hybridoma growth factor; and plasmacytoma growthfactor) is a multifunctional cytokine involved in numerous biologicalprocesses such as the regulation of the acute inflammatory response, themodulation of specific immune responses including B- and T-celldifferentiation, bone metabolism, thrombopoiesis, epidermalproliferation, menses, neuronal cell differentiation, neuroprotection,aging, cancer, and the inflammatory reaction occurring in Alzheimer'sdisease. See A. Papassotiropoulos et al, Neurobiology of Aging,22:863-871 (2001).

IL-6 is a member of a family of cytokines that promote cellularresponses through a receptor complex consisting of at least one subunitof the signal-transducing glycoprotein gp130 and the IL-6 receptor(“IL-6R”) (also known as gp80). The IL-6R may also be present in asoluble form (“sIL-6R”). IL-6 binds to IL-6R, which then dimerizes thesignal-transducing receptor gp130. See Jones, S A, J. Immunology,175:3463-3468 (2005).

In humans, the gene encoding IL-6 is organized in five exons and fourintrons, and maps to the short arm of chromosome 7 at 7p21. Translationof IL-6 RNA and post-translational processing result in the formation ofa 21 to 28 kDa protein with 184 amino acids in its mature form. See A.Papassotiropoulos, et al, Neurobiology of Aging, 22:863-871 (2001).

As set forth in greater detail herein IL-6 is believed to play a role inthe development of a multitude of diseases and disorders, including butnot limited to fatigue, cachexia, autoimmune diseases, diseases of theskeletal system, cancer, heart disease, obesity, diabetes, asthma,alzheimer's disease and multiple sclerosis. Due to the perceivedinvolvement of IL-6 in a wide range of diseases and disorders, thereremains a need in the art for compositions and methods useful forpreventing or treating diseases associated with IL-6, as well as methodsof screening to identify patients having diseases or disordersassociated with IL-6. Particularly preferred anti-IL-6 compositions arethose having minimal or minimizing adverse reactions when administeredto the patient. Compositions or methods that reduce or inhibit diseasesor disorders associated with IL-6 are beneficial to the patient in needthereof.

The function of IL-6 is not restricted to the immune response as it actsin hematopoiesis, thrombopoiesis, osteoclast formation, elicitation ofhepatic acute phase response resulting in the elevation of C-reactiveprotein (CRP) and serum amyloid A (SAA) protein. It is known to be agrowth factor for epidermal keratinocytes, renal mesangial cells,myeloma and plasmacytoma cells (Grossman et al., 1989 Prot Natl AcadSci., 86, (16) 6367-6371; Horii et al., 1989, J Immunol, 143, 12,3949-3955; Kawano et al., 1988, Nature 332, 6159, 83-85). IL-6 isproduced by a wide range of cell types including monocytes/macrophages,fibroblasts, epidermal keratinocytes, vascular endothelial cells, renalmessangial cells, glial cells, condrocytes, T and B-cells and sometumour cells (Akira et al, 1990, FASEB J., 4, 11, 2860-2867). Except fortumour cells that constitutively produce IL-6, normal cells do notexpress IL-6 unless appropriately stimulated.

Elevated IL-6 levels have been observed in many types of cancer,including breast cancer, leukemia, ovarian cancer, prostate cancer,pancreatic cancer, lymphoma, lung cancer, renal cell carcinoma,colorectal cancer, and multiple myeloma (e.g., Chopra et al., 2004,MJAFI 60:45-49; Songur et al., 2004, Tumori 90:196-200; Blay et al.,1992, Cancer Research 52:3317-3322; Nikiteas et al., 2005, World J.Gasterenterol. 11:1639-1643; reviewed in Heikkila et al., 2008, Eur JCancer, 44:937-945). As noted above, IL-6 is known or suspected to playa role in promoting proliferation or survival of at least some types ofcancer. Moreover, some of these studies have demonstrated correlationbetween IL-6 levels and patient oucome. Together, these results suggestthe possibility that inhibition of IL-6 can be therapeuticallybeneficial. Indeed, clinical studies (reviewed in Trikha et al., 2003,Clinical Cancer Research 9:4653-4665) have shown some improvement inpatient outcomes due to administration of various anti-IL-6 antibodies,particularly in those cancers in which IL-6 plays a direct rolepromoting cancer cell proliferation or survival.

As noted above, IL-6 stimulates the hepatic acute phase response,resulting in increased production of CRP and elevated serum CRP levels.For this reason, C-reactive protein (CRP) has been reported to comprisea surrogate marker of IL-6 activity. Thus, elevated IL-6 activity can bedetected through measurement of serum CRP. Conversely, effectivesuppression of IL-6 activity, e.g., through administration of aneutralizing anti-IL-6 antibody, can be detected by the resultingdecrease in serum CRP levels.

A recent clinical trial demonstrated that administration of rosuvastatinto apparently healthy individuals having elevated CRP (greater than 2.0mg/1) reduced their CRP levels by 37% and greatly decreased theincidence of myocardial infarction, stroke, arterial revascularization,hospitalization for unstable angina, or death from cardiovascularcauses. Ridker et al., N Engl J Med. 2008 Nov. 9 [Epub ahead of print].

In addition to its direct role in pathogenesis of some cancers and otherdiseases, chronically elevated IL-6 levels appear to adversely affectpatient well-being and quality of life. For example, elevated IL-6levels have been reported to be associated with cachexia and fever, andreduced serum albumin. Gauldie et al., 1987, PNAS 84:7251-7253; Heinricet al., 1990, 265:621-636; Zamir et al., 1993, Metabolism 42:204-208;Zamir et al., 1992, Arch Surg, 127:170-174. Inhibition of IL-6 by aneutralizing antibody has been reported to ameliorate fever and cachexiain cancer patients, though improvement in these patients' serum albuminlevel has not been reported (Emille et al., 1994, Blood, 84:2472-2479;Blay et al., 1992, Cancer Research 52:3317-3322; Bataille et al., 1995,Blood, 86: 685-691).

Numerous studies have suggested that CRP is a valuable prognostic factorin cancer patients, with elevated CRP levels predicting poor outcome.See, e.g., Hefler et al, Clin Cancer Res, 2008 Feb. 1; 14(3):710-4;Nagaoka et al, Liver Int, 2007 October; 27(8):1091-7; Heikkila et al, JEpidemiol Community Health, 2007 September; 61(9):824-33, Review; Haraet al, Anticancer Res, 2007 July-August; 27(4C):3001-4; Polterauer etal, Gynecol Oncol, 2007 October; 107(1):114-7, Epub 2007 Jul. 6;Tingstedt et al, Scand J Gastroenterol, 2007 June; 42(6):754-9; Suh etal, Support Care Cancer, 2007 June; 15(6):613-20, Epub 2007 Jan. 18;Gerhardt et al, World J Gastroenterol, 2006 Sep. 14; 12(34):5495-500;McArdle et al, Urol Int, 2006; 77(2):127-9; Guillem et al, DisEsophagus, 2005; 18(3):146-50; Brown et al, Cancer, 2005 Jan. 15;103(2):377-82. Decreased serum albumin (hypoalbuminemia) is alsoassociated with increased morbidity and mortality in many criticalillnesses, including cancers (e.g., Vigano et al., Arch Intern Med, 2000Mar. 27; 160(6):861-8; Hauser et al., Support Care Cancer, 2006 October;14(10):999-1011; Seve et al., Cancer, 2006 Dec. 1; 107(11):2698-705).The apparent link between hypoalbuminemia and poor patient oucomesuggests that restoring albumin levels through direct albumin infusioncould promote patient survival, however, albumin infusion has notimproved survival of patients with advanced cancer (Demirkazik et al.,Proc Am Soc Clin Oncol 21: 2002 (abstr 2892)) or other critically illpatients groups (reviewed in Wilkes et al., Ann Intern Med, 2001 Aug. 7;135(3):149-64).

The Glasgow Prognostic Score (GPS) is an inflammation-based prognosticscore that combines levels of albumin (<35 mg/L=1 point) and CRP (>10mg/L=1 point) (Forrest et al., Br J Cancer, 2004 May 4; 90(9):1704-6).Since its introduction in 2004, the Glasgow Prognostic Score has alreadybeen shown to have prognostic value as a predictor of mortality innumerous cancers, including gastro-oesophageal cancer, non-small-celllung cancer, colorectal cancer, breast cancer, ovarian cancer,bronchogenic cancer, and metastatic renal cancer (Forrest et al., Br JCancer, 2004 May 4; 90(9):1704-6; Sharma et al., Clin Colorectal Cancer,2008 September; 7(5):331-7; Sharma et al., Eur J Cancer, 2008 January;44(2):251-6; McMillan et al., Nutr Cancer, 2001; 41(1-2):64-9; McMillan,Proc Nutr Soc, 2008 August; 67(3):257-62; Ramsey et al., Cancer, 2007Jan. 15; 109(2):205-12). Because the combination of elevated CRP andreduced albumin predicts cancer patient mortality, a treatment that bothlowers CRP and raises albumin would suggest a strong possiblity of alsopromoting patient survival.

U.S. patent application publication no. 20080081041 (relating totreatment of cancer using an anti-IL-6 antibody) discloses that sinceIL-6 is associated with disease activity and since CRP is a surrogatemarker of IL-6 activity, sustained suppression of CRP by neutralizationof IL-6 by their anti-IL-6 antibody (CNTO 328, Zaki et al., Int JCancer, 2004 Sep. 10; 111(4):592-5) may be assumed necessary to achievebiological activity. The same patent application indicates that therelationship between IL-6 and CRP in patients with benign and malignantprostate disease was previously examined by McArdle (McArdle et al. 2004Br J Cancer 91(10):1755-1757). McArdle reportedly found no significantdifferences between the concentrations of IL-6 and CRP in the patientswith benign disease compared with prostate cancer patients, in thecancer patients there was a significant increase in both IL-6 and CRPconcentration with increasing tumor grade. The median serum CRP valuefor the 86 subjects with prostate cancer was 1.8 mg/L. Based thereoonthe inventors in this patent application postulate a proposed dose andschedule wherein 6 mg/kg of an anti-IL-6 antibody (CNTO 328) isadministered every 2 weeks and allege that this is likely to achievesustained suppression of CRP in subjects with metastatic HRPC.

IL-6 signaling is mediated by the Jak-Tyk family of cytoplasmic tyrosinekinases, including JAK1, JAK2, and JAK3 (reviewed in Murray J Immunol.2007 Mar. 1; 178(5):2623-9). Sivash et al. report abrogation ofIL-6-mediated JAK signalling by the cyclopentenone prostaglandin15d-PGD₂ in oral squamous carcinoma cells. British Journal of Cancer(2004) 91, 1074-1080. These results suggest that inhibitors of JAK1,JAK2, or JAK3 could be employed as antagonists of IL-6.

Ulanova et al. report that inhibition of the nonreceptor proteintyrosine kinase Syk (using siRNA) decreased production of IL-6 byepithelial cells. Am J Physiol Lung Cell Mol Physiol. 2005 March;288(3):L497-507. These results suggest that an inhibitor of Syk could beemployed as an antagonist of IL-6.

Kedar et al. report that treatment with thalidomide significantlyreduced serum levels of CRP and IL-6 to normal or near normal levels ina substantial fraction of renal cell carcinoma patients. Int J Cancer.2004 Jun. 10; 110(2):260-5. These results suggest that thalidomide, andpossibly derivatives thereof, such as lenalidomide, may be usefulantagonists of IL-6.

In addition, another published patent application, US 20070292420teaches a Phase I dose escalating study using an anti-IL-6 (cCLB-8)antibody for treating refractory patients with advanced stage multiplemyeloma (N=12) and indicate that this study demonstrated that somepatients had disease stabilization. The application also reports thatafter discontinuation of treatment there was acceleration in theincrease of M protein levels, suggesting disease re-bound after thewithdrawal of therapy. Anti-IL-6 cCLB-8 antibody inhibited freecirculating IL-6.

The application also indicates that this antibody trial resulted in notoxicity (except transient thrombocytopenia in two heavily pretreatedpatients) or allergic reactions were observed and that C-reactiveprotein (CRP) decreased below detection level in all patients. Theirantibody (cCLB-8 antibody) reportedly possessed a circulating half-lifeof 17.8 days, and that there was no human anti-chimeric antibody (HACA)immune response observed (van Zaanen et al. 1998). They allege that theadministration of CNTO 328 did not cause changes in blood pressure,pulse rate, temperature, hemoglobin, liver functions and renalfunctions. Except for transient thrombocytopenia in two heavilypretreated patients, no toxicity or allergic reactions allegedly wereobserved, and there was no human anti-chimeric antibody (HACA) immuneresponse observed. Three patients in their study reportedly developedinfection-related complications during therapy, however, a possiblerelation with anti-IL-6 cCLB-8 antibody was concluded by the inventorsto be unlikely because infectious complications are reportedly common inend stage multiple myeloma and are a major cause of death. They concludebased on their results that this anti-IL-6 cCLB-8 antibody was safe inmultiple myeloma patients.

BRIEF SUMMARY OF THE INVENTION

The present invention is an extension of Applicants' previous inventionwhich is directed to specific antibodies and fragments thereof havingbinding specificity for IL-6, in particular antibodies having specificepitopic specificity and/or functional properties and novel therapiesusing these and other anti-IL-6 antibodies. One embodiment of theinvention encompasses specific humanized antibodies and fragmentsthereof capable of binding to IL-6 and/or the IL-6/IL-6R complex. Theseantibodies may bind soluble IL-6 or cell surface expressed IL-6. Also,these antibodies may inhibit the formation or the biological effects ofone or more of IL-6, IL-6/IL-6R complexes, IL-6/IL-6R/gp130 complexesand/or multimers of IL-6/IL-6R/gp130. The present invention relates tonovel therapies and therapeutic protocols using anti-IL-6 antibodies,preferably those described herein. In particular, the present inventionpertains to methods of preventing or treating thrombosis in a patient inneed thereof, e.g., a patient showing elevated D-dimer and/or CRP levelsprior to treatment, comprising administering to the patient an IL-6antagonist, such as those identified infra, e.g., an anti-IL-6 antibodyor antibody fragment, whereby the patient's coagulation profile isimproved or restored to a normal condition. In some embodiments thesemethods may further include the administration of other actives such asstatins that may further help (synergize) with the IL-6 antagonist andthereby more effectively treat or prevent thrombosis.

In a preferred embodiment this is effected by the administration of theantibodies described herein, comprising the sequences of the V_(H),V_(L) and CDR polypeptides described herein, and the polynucleotidesencoding them. In more specific embodiments of the invention theseantibodies will block gpI30 activation and/or possess binding affinities(Kds) less than 50 picomolar and/or K_(off) values less than or equal to10⁻⁴S⁻¹.

In another embodiment of the invention these antibodies and humanizedversions will be derived from rabbit immune cells (B lymphocytes) andmay be selected based on their homology (sequence identity) to humangerm line sequences. These antibodies may require minimal or no sequencemodifications, thereby facilitating retention of functional propertiesafter humanization.

In another embodiment of the invention the subject antibodies may beselected based on their activity in functional assays such as IL-6driven T1165 proliferation assays, IL-6 simulated HepG2 haptoglobinproduction assays, and the like. A further embodiment of the inventionis directed to fragments from anti-IL-6 antibodies encompassing V_(H),V_(L) and CDR polypeptides, e.g., derived from rabbit immune cells andthe polynucleotides encoding the same, as well as the use of theseantibody fragments and the polynucleotides encoding them in the creationof novel antibodies and polypeptide compositions capable of recognizingIL-6 and/or IL-6/IL-6R complexes or IL-6/IL-6R/gpI30 complexes and/ormultimers thereof.

The invention also contemplates the administration of conjugates ofanti-IL-6 antibodies and binding fragments thereof conjugated to one ormore functional or detectable moieties. The invention also contemplatesmethods of making said humanized anti-IL-6 or anti-IL-6/IL-6R complexantibodies and binding fragments thereof. In one embodiment, bindingfragments include, but are not limited to, Fab, Fab′, F(ab)₂, Fv andscFv fragments.

Embodiments of the invention pertain to the use of anti-IL-6 antibodiesfor the diagnosis, assessment and treatment of diseases and disordersassociated with IL-6 or aberrant expression thereof. The invention alsocontemplates the use of fragments of anti-IL-6 antibodies for thediagnosis, assessment and treatment of diseases and disorders associatedwith IL-6 or aberrant expression thereof. Preferred usages of thesubject antibodies are the treatment and prevention of cancer associatedfatigue, and/or cachexia and rheumatoid arthritis.

Other embodiments of the invention relate to the production of anti-IL-6antibodies in recombinant host cells, preferably diploid yeast such asdiploid Pichia and other yeast strains.

Another embodiment of the invention relates to methods of improvingsurvivability or quality of life of a patient diagnosed with cancer,comprising administering to the patient an anti-IL-6 antibody orantibody fragment, whereby the patient's serum C-reactive protein(“CRP”) level is reduced, and monitoring the patient to assess thereduction in the patient's serum CRP level, wherein the anti-IL-6antibody or antibody fragment may specifically bind to the same linearor conformational epitope(s) and/or compete for binding to the samelinear or conformational epitope(s) on an intact human IL-6 polypeptideor antibody fragment thereof as an anti-IL-6 antibody selected from thegroup consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10,Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22,Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34,Ab35, and Ab36.

Another embodiment of the invention relates to methods of improvingmuscular strength in a patient diagnosed with cancer, comprisingadministering to the patient an anti-IL-6 antibody or antibody fragment,whereby the patient's muscular strength is improved, and monitoring thepatient to assess muscular strength, wherein the anti-IL-6 antibody orantibody fragment may specifically bind to the same linear orconformational epitope(s) and/or compete for binding to the same linearor conformational epitope(s) on an intact human IL-6 polypeptide orantibody fragment thereof as an anti-IL-6 antibody selected from thegroup consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10,Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22,Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34,Ab35, and Ab36. In such methods preferably the patient's muscularstrength is improved by at least about 15% within approximately 4 weeksof administering the anti-IL-6 antibody or antibody fragment, asmeasured by the Hand Grip Strength test and more preferably thepatient's muscular strength is improved by at least about 20% withinapproximately 4 weeks of administering the anti-IL-6 antibody orantibody fragment, as measured by the Hand Grip Strength test.

Another embodiment of the invention relates to methods of increasingserum albumin in a patient in need thereof, comprising administering tothe patient an anti-IL-6 antibody or antibody fragment, whereby thepatient's serum albumin level is improved, and monitoring the patient toassess serum albumin level, wherein the anti-IL-6 antibody or antibodyfragment may specifically bind to the same linear or conformationalepitope(s) and/or compete for binding to the same linear orconformational epitope(s) on an intact human IL-6 polypeptide orantibody fragment thereof as an anti-IL-6 antibody selected from thegroup consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10,Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22,Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34,Ab35, and Ab36. Preferably, these methods are effected under conditionswhereby the patient's survivability is improved, and/or under conditionswherein the serum albumin level is increased by about 5 g/L withinapproximately 6 weeks of administering the anti-IL-6 antibody orantibody fragment. These patients will include, without limitationthereto, those diagnosed with rheumatoid arthritis, cancer, advancedcancer, liver disease, renal disease, inflammatory bowel disease,celiac's disease, trauma, burns, other diseases associated with reducedserum albumin, or any combination thereof.

Another embodiment of the invention relates to methods of preventing ortreating thrombosis in a patient in a state of hypercoagulation,comprising administering to the patient an IL-6 antagonist, e.g. ananti-IL-6 antibody or antibody fragment, whereby the patient'scoagulation profile is improved or restored to a normal condition, andmonitoring the patient to assess coagulation profile. In such methods ifthe IL-6 antagonist is an anti-IL-6 antibody or antibody fragmentpreferably this antibody may specifically bind to the same linear orconformational epitope(s) and/or compete for binding to the same linearor conformational epitope(s) on an intact human IL-6 polypeptide orantibody fragment thereof as an anti-IL-6 antibody selected from thegroup consisting of AbI, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, AbI0,AbI1, AbI2, AbI3, AbI4, AbI5, AbI6, AbI7, AbI8, AbI9, Ab20, Ab2I, Ab22,Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29, Ab30, Ab3I, Ab32, Ab33, Ab34,Ab35, and Ab36. In the inventive methods of preventing or treatingthrombosis, the patient's coagulation profile is assessed by measurementof the patient's serum level of one or more of D-dimer, Factor II,Factor V, Factor VIII, Factor IX, Factor XI, Factor XII, F/fibrindegradation products, thrombin-antithrombin III complex, fibrinogen,plasminogen, prothrombin, and von Willebrand factor and preferably by amethod including measuring the patient's serum D-dimer level prior toadministration of the anti-IL-6 antibody, and administering theanti-IL-6 antibody or antibody fragment if the patient's serum D-dimerlevel is elevated. In addition, the levels of C reactive protein mayalso be asessed in the patient prior to treatment and if elevated thismay be used as a further indicator as to an increased risk of thrombosisin the patient.

An embodiment of the invention relates to methods of treating a patienthaving a disease or condition associated with hypercoagulation, whichmay comprise administering to the patient an IL-6 antagonist, wherebythe patient's coagulation profile is improved or restored to normal, andmonitoring the patient to assess coagulation profile.

In an embodiment of the invention, the patient may have elevated serumD-dimer levels prior to treatment.

In an embodiment of the invention, the patient may have elevatedC-reactive protein (CRP) levels prior to treatment.

In an embodiment of the invention, the method may further comprise theadministration of at least one statin.

In an embodiment of the invention, the IL-6 antagonist may target IL-6,IL-6 receptor alpha, gp130, p38 MAP kinase, JAK1, JAK2, JAK3, SYK, orany combination thereof.

In an embodiment of the invention, the IL-6 antagonist may comprise anantibody, an antibody fragment, a peptide, a glycoalkoid, an antisensenucleic acid, a ribozyme, a retinoid, an avemir, a small molecule, orany combination thereof.

In an embodiment of the invention, the IL-6 antagonist may comprise ananti-IL-6R, anti-gp130, anti-p38 MAP kinase, anti-JAK1, anti-JAK2,anti-JAK3, or anti-SYK antibody or antibody fragment.

In one embodiment of the invention, the IL-6 antagonist may comprise asmall molecule comprising thalidomide, lenalidomide, or any combinationthereof.

In an embodiment of the invention, the antagonist may comprise ananti-IL-6 antibody or antibody fragment.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may specifically bind to the same linear or conformationalepitope(s) and/or compete for binding to the same linear orconformational epitope(s) on an intact human IL-6 polypeptide orantibody fragment thereof as an anti-IL-6 antibody selected from thegroup consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10,Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22,Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34,Ab35, and Ab36.

In an embodiment of the invention, the anti-IL-6 antibody may bind tothe same linear or conformational epitope(s) and/or compete for bindingto the same linear or conformational epitope(s) on an intact human IL-6polypeptide or a fragment thereof as Ab1.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may specifically bind to the same linear or conformationalepitope(s) on an intact human IL-6 polypeptide or antibody fragmentthereof as an anti-IL-6 antibody selected from the group consisting ofAb1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13,Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25,Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, and Ab36.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may specifically bind to the same linear or conformationalepitope(s) on an intact human IL-6 polypeptide or a fragment thereof asAb1.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may specifically bind to the same linear or conformationalepitopes on an intact IL-6 polypeptide or antibody fragment thereof thatis (are) specifically bound by Ab1 and wherein said epitope(s) whenascertained by epitopic mapping using overlapping linear peptidefragments which span the full length of the native human IL-6polypeptide includes one or more residues comprised in IL-6 fragmentsselected from those respectively encompassing amino acid residues 37-51,amino acid residues 70-84, amino acid residues 169-183, amino acidresidues 31-45 and/or amino acid residues 58-72.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may comprise at least 2 complementarity determining regions(CDRs) in each the variable light and the variable heavy regions whichare identical to those contained in an anti-IL-6 antibody selected fromthe group consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9,Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21,Ab22, Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33,Ab34, Ab35, and Ab36.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may comprise at least 2 complementarity determining regions(CDRs) in each the variable light and the variable heavy regions whichare identical to those contained in Ab1.

In an embodiment of the invention, all of the CDRs in the anti-IL-6antibody or antibody fragment may be identical to the CDRs contained inan anti-IL-6 antibody selected from the group consisting of Ab1, Ab2,Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15,Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25, Ab26, Ab27,Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, and Ab36.

In an embodiment of the invention, all of the CDRs in the anti-IL-6antibody or antibody fragment may be identical to the CDRs contained inAb1.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may be aglycosylated.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may contain an Fc region that has been modified to altereffector function, half-life, proteolysis, and/or glycosylation.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may be a human, humanized, single chain or chimeric antibody.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may be a humanized antibody derived from a rabbit (parent)anti-IL-6 antibody.

In an embodiment of the invention, the framework regions (FRs) in thevariable light region and the variable heavy regions of said anti-IL-6antibody or antibody fragment respectively may be human FRs which areunmodified or which have been modified by the substitution of at most 2or 3 human FR residues in the variable light or heavy chain region withthe corresponding FR residues of the parent rabbit antibody, and thehuman FRs may have been derived from human variable heavy and lightchain antibody sequences which have been selected from a library ofhuman germline antibody sequences based on their high level of homologyto the corresponding rabbit variable heavy or light chain regionsrelative to other human germline antibody sequences contained in thelibrary.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may be administered to the patient with a frequency at mostonce per period of approximately four weeks, approximately eight weeks,approximately twelve weeks, approximately sixteen weeks, approximatelytwenty weeks, or approximately twenty-four weeks.

In an embodiment of the invention, the patient's coagulation profile mayremain improved for an entire period intervening two consecutiveanti-IL-6 antibody administrations.

In an embodiment of the invention, the patient may have been diagnosedwith cancer selected from Acanthoma, Acinic cell carcinoma, Acousticneuroma, Acral lentiginous melanoma, Acrospiroma, Acute eosinophilicleukemia, Acute lymphoblastic leukemia, Acute megakaryoblastic leukemia,Acute monocytic leukemia, Acute myeloblastic leukemia with maturation,Acute myeloid dendritic cell leukemia, Acute myeloid leukemia, Acutepromyelocytic leukemia, Adamantinoma, Adenocarcinoma, Adenoid cysticcarcinoma, Adenoma, Adenomatoid odontogenic tumor, Adrenocorticalcarcinoma, Adult T-cell leukemia, Aggressive NK-cell leukemia,AIDS-Related Cancers, AIDS-related lymphoma, Alveolar soft part sarcoma,Ameloblastic fibroma, Anal cancer, Anaplastic large cell lymphoma,Anaplastic thyroid cancer, Angioimmunoblastic T-cell lymphoma,Angiomyolipoma, Angiosarcoma, Appendix cancer, Astrocytoma, Atypicalteratoid rhabdoid tumor, Basal cell carcinoma, Basal-like carcinoma,B-cell leukemia, B-cell lymphoma, Bellini duct carcinoma, Biliary tractcancer, Bladder cancer, Blastoma, Bone Cancer, Bone tumor, Brain StemGlioma, Brain Tumor, Breast Cancer, Brenner tumor, Bronchial Tumor,Bronchioloalveolar carcinoma, Brown tumor, Burkitt's lymphoma, Cancer ofUnknown Primary Site, Carcinoid Tumor, Carcinoma, Carcinoma in situ,Carcinoma of the penis, Carcinoma of Unknown Primary Site,Carcinosarcoma, Castleman's Disease, Central Nervous System EmbryonalTumor, Cerebellar Astrocytoma, Cerebral Astrocytoma, Cervical Cancer,Cholangiocarcinoma, Chondroma, Chondrosarcoma, Chordoma,Choriocarcinoma, Choroid plexus papilloma, Chronic Lymphocytic Leukemia,Chronic monocytic leukemia, Chronic myelogenous leukemia, ChronicMyeloproliferative Disorder, Chronic neutrophilic leukemia, Clear-celltumor, Colon Cancer, Colorectal cancer, Craniopharyngioma, CutaneousT-cell lymphoma, Degos disease, Dermatofibrosarcoma protuberans, Dermoidcyst, Desmoplastic small round cell tumor, Diffuse large B celllymphoma, Dysembryoplastic neuroepithelial tumor, Embryonal carcinoma,Endodermal sinus tumor, Endometrial cancer, Endometrial Uterine Cancer,Endometrioid tumor, Enteropathy-associated T-cell lymphoma,Ependymoblastoma, Ependymoma, Epithelioid sarcoma, Erythroleukemia,Esophageal cancer, Esthesioneuroblastoma, Ewing Family of Tumor, EwingFamily Sarcoma, Ewing's sarcoma, Extracranial Germ Cell Tumor,Extragonadal Germ Cell Tumor, Extrahepatic Bile Duct Cancer,Extramammary Paget's disease, Fallopian tube cancer, Fetus in fetu,Fibroma, Fibrosarcoma, Follicular lymphoma, Follicular thyroid cancer,Gallbladder Cancer, Gallbladder cancer, Ganglioglioma, Ganglioneuroma,Gastric Cancer, Gastric lymphoma, Gastrointestinal cancer,Gastrointestinal Carcinoid Tumor, Gastrointestinal Stromal Tumor,Gastrointestinal stromal tumor, Germ cell tumor, Germinoma, Gestationalchoriocarcinoma, Gestational Trophoblastic Tumor, Giant cell tumor ofbone, Glioblastoma multiforme, Glioma, Gliomatosis cerebri, Glomustumor, Glucagonoma, Gonadoblastoma, Granulosa cell tumor, Hairy CellLeukemia, Hairy cell leukemia, Head and Neck Cancer, Head and neckcancer, Heart cancer, Hemangioblastoma, Hemangiopericytoma,Hemangiosarcoma, Hematological malignancy, Hepatocellular carcinoma,Hepatosplenic T-cell lymphoma, Hereditary breast-ovarian cancersyndrome, Hodgkin Lymphoma, Hodgkin's lymphoma, Hypopharyngeal Cancer,Hypothalamic Glioma, Inflammatory breast cancer, Intraocular Melanoma,Islet cell carcinoma, Islet Cell Tumor, Juvenile myelomonocyticleukemia, Kaposi Sarcoma, Kaposi's sarcoma, Kidney Cancer, Klatskintumor, Krukenberg tumor, Laryngeal Cancer, Laryngeal cancer, Lentigomaligna melanoma, Leukemia, Leukemia, Lip and Oral Cavity Cancer,Liposarcoma, Lung cancer, Luteoma, Lymphangioma, Lymphangiosarcoma,Lymphoepithelioma, Lymphoid leukemia, Lymphoma, Macroglobulinemia,Malignant Fibrous Histiocytoma, Malignant fibrous histiocytoma,Malignant Fibrous Histiocytoma of Bone, Malignant Glioma, MalignantMesothelioma, Malignant peripheral nerve sheath tumor, Malignantrhabdoid tumor, Malignant triton tumor, MALT lymphoma, Mantle celllymphoma, Mast cell leukemia, Mediastinal germ cell tumor, Mediastinaltumor, Medullary thyroid cancer, Medulloblastoma, Medulloblastoma,Medulloepithelioma, Melanoma, Melanoma, Meningioma, Merkel CellCarcinoma, Mesothelioma, Mesothelioma, Metastatic Squamous Neck Cancerwith Occult Primary, Metastatic urothelial carcinoma, Mixed Mülleriantumor, Monocytic leukemia, Mouth Cancer, Mucinous tumor, MultipleEndocrine Neoplasia Syndrome, Multiple Myeloma, Multiple myeloma,Mycosis Fungoides, Mycosis fungoides, Myelodysplastic Disease,Myelodysplastic Syndromes, Myeloid leukemia, Myeloid sarcoma,Myeloproliferative Disease, Myxoma, Nasal Cavity Cancer, NasopharyngealCancer, Nasopharyngeal carcinoma, Neoplasm, Neurinoma, Neuroblastoma,Neuroblastoma, Neurofibroma, Neuroma, Nodular melanoma, Non-HodgkinLymphoma, Non-Hodgkin lymphoma, Nonmelanoma Skin Cancer, Non-Small CellLung Cancer, Ocular oncology, Oligoastrocytoma, Oligodendroglioma,Oncocytoma, Optic nerve sheath meningioma, Oral Cancer, Oral cancer,Oropharyngeal Cancer, Osteosarcoma, Osteosarcoma, Ovarian Cancer,Ovarian cancer, Ovarian Epithelial Cancer, Ovarian Germ Cell Tumor,Ovarian Low Malignant Potential Tumor, Paget's disease of the breast,Pancoast tumor, Pancreatic Cancer, Pancreatic cancer, Papillary thyroidcancer, Papillomatosis, Paraganglioma, Paranasal Sinus Cancer,Parathyroid Cancer, Penile Cancer, Perivascular epithelioid cell tumor,Pharyngeal Cancer, Pheochromocytoma, Pineal Parenchymal Tumor ofIntermediate Differentiation, Pineoblastoma, Pituicytoma, Pituitaryadenoma, Pituitary tumor, Plasma Cell Neoplasm, Pleuropulmonaryblastoma, Polyembryoma, Precursor T-lymphoblastic lymphoma, Primarycentral nervous system lymphoma, Primary effusion lymphoma, PrimaryHepatocellular Cancer, Primary Liver Cancer, Primary peritoneal cancer,Primitive neuroectodermal tumor, Prostate cancer, Pseudomyxomaperitonei, Rectal Cancer, Renal cell carcinoma, Respiratory TractCarcinoma Involving the NUT Gene on Chromosome 15, Retinoblastoma,Rhabdomyoma, Rhabdomyosarcoma, Richter's transformation, Sacrococcygealteratoma, Salivary Gland Cancer, Sarcoma, Schwannomatosis, Sebaceousgland carcinoma, Secondary neoplasm, Seminoma, Serous tumor,Sertoli-Leydig cell tumor, Sex cord-stromal tumor, Sézary Syndrome,Signet ring cell carcinoma, Skin Cancer, Small blue round cell tumor,Small cell carcinoma, Small Cell Lung Cancer, Small cell lymphoma, Smallintestine cancer, Soft tissue sarcoma, Somatostatinoma, Soot wart,Spinal Cord Tumor, Spinal tumor, Splenic marginal zone lymphoma,Squamous cell carcinoma, Stomach cancer, Superficial spreading melanoma,Supratentorial Primitive Neuroectodermal Tumor, Surfaceepithelial-stromal tumor, Synovial sarcoma, T-cell acute lymphoblasticleukemia, T-cell large granular lymphocyte leukemia, T-cell leukemia,T-cell lymphoma, T-cell prolymphocytic leukemia, Teratoma, Terminallymphatic cancer, Testicular cancer, Thecoma, Throat Cancer, ThymicCarcinoma, Thymoma, Thyroid cancer, Transitional Cell Cancer of RenalPelvis and Ureter, Transitional cell carcinoma, Urachal cancer, Urethralcancer, Urogenital neoplasm, Uterine sarcoma, Uveal melanoma, VaginalCancer, Verner Morrison syndrome, Verrucous carcinoma, Visual PathwayGlioma, Vulvar Cancer, Waldenström's macroglobulinemia, Warthin's tumor,Wilms' tumor, or any combination thereof.

In an embodiment of the invention, the patient may have been diagnosedwith a cancer selected from Colorectal Cancer, Non-Small Cell LungCancer, Cholangiocarcinoma, Mesothelioma, Castleman's disease, RenalCell Carcinoma, or any combination thereof.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may comprise a VH polypeptide sequence selected from the groupconsisting of: SEQ ID NO: 3, 18, 19, 22, 38, 54, 70, 86, 102, 117, 118,123, 139, 155, 171, 187, 203, 219, 235, 251, 267, 283, 299, 315, 331,347, 363, 379, 395, 411, 427, 443, 459, 475, 491, 507, 523, 539, 555 andSEQ ID NO: 571; and may further comprise a VL polypeptide sequenceselected from the group consisting of: SEQ ID NO: 2, 20, 21, 37, 53, 69,85, 101, 119, 122, 138, 154, 170, 186, 202, 218, 234, 250, 266, 282,298, 314, 330, 346, 362, 378, 394, 410, 426, 442, 458, 474, 490, 506,522, 538, 554 and SEQ ID NO: 570 or a variant thereof wherein one ormore of the framework residues (FR residues) in said VH or VLpolypeptide may have been substituted with another amino acid residueresulting in an anti-IL-6 antibody or antibody fragment thatspecifically binds human IL-6.

In an embodiment of the invention, one or more of said FR residues maybe substituted with an amino acid present at the corresponding site in aparent rabbit anti-IL-6 antibody from which the complementaritydetermining regions (CDRs) contained in said VH or VL polypeptides havebeen derived or by a conservative amino acid substitution.

In an embodiment of the invention, said anti-IL-6 antibody or antibodyfragment may be humanized.

In an embodiment of the invention, said anti-IL-6 antibody or antibodyfragment may be chimeric.

In an embodiment of the invention, said anti-IL-6 antibody or antibodyfragment further may comprise a human Fc.

In an embodiment of the invention, said human Fc may be derived fromIgG1, IgG2, IgG3, IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10, IgG11,IgG12, IgG13, IgG14, IgG15, IgG16, IgG17, IgG18 or IgG19.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may comprise a polypeptide having at least 90% sequencehomology to one or more of the polypeptide sequences of SEQ ID NO: 3,18, 19, 22, 38, 54, 70, 86, 102, 117, 118, 123, 139, 155, 171, 187, 203,219, 235, 251, 267, 283, 299, 315, 331, 347, 363, 379, 395, 411, 427,443, 459, 475, 491, 507, 523, 539, 555, 571, 2, 20, 21, 37, 53, 69, 85,101, 119, 122, 138, 154, 170, 186, 202, 218, 234, 250, 266, 282, 298,314, 330, 346, 362, 378, 394, 410, 426, 442, 458, 474, 490, 506, 522,538, 554, and 570.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may have an elimination half-life of at least about 22 days, atleast about 25 days, or at least about 30 days.

In an embodiment of the invention, the IL-6 antagonist may beco-administered with a chemotherapy agent, including without limitationthereto: VEGF antagonists, EGFR antagonists, platins, taxols,irinotecan, 5-fluorouracil, gemcytabine, leucovorine, steroids,cyclophosphamide, melphalan, vinca alkaloids (e.g., vinblastine,vincristine, vindesine and vinorelbine), mustines, tyrosine kinaseinhibitors, radiotherapy, sex hormone antagonists, selective androgenreceptor modulators, selective estrogen receptor modulators, PDGFantagonists, TNF antagonists, IL-1 antagonists, interleukins (e.g. IL-12or IL-2), IL-12R antagonists, Toxin conjugated monoclonal antibodies,tumor antigen specific monoclonal antibodies, Erbitux, Avastin,Pertuzumab, anti-CD20 antibodies, Rituxan, ocrelizumab, ofatumumab,DXL625, herceptin, or any combination thereof.

In an embodiment of the invention, the another therapeutic compound maybe a statin.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may be directly or indirectly attached to a detectable label ortherapeutic agent.

In an embodiment of the invention, the anti-IL-6 antibody or antibodyfragment may be Ab1 or a fragment thereof.

In an embodiment of the invention, the disease or condition may beselected from acute venous thrombosis, pulmonary embolism, thrombosisduring pregnancy, hemorrhagic skin necrosis, acute or chronicdisseminated intravascular coagulation (DIC), clot formation fromsurgery, long bed rest, long periods of immobilization, venousthrombosis, fulminant meningococcemia, acute thrombotic stroke, acutecoronary occlusion, acute peripheral arterial occlusion, massivepulmonary embolism, axillary vein thrombosis, massive iliofemoral veinthrombosis, occluded arterial cannulae, occluded venous cannulae,cardiomyopathy, venoocclusive disease of the liver, hypotension,decreased cardiac output, decreased vascular resistance, pulmonaryhypertension, diminished lung compliance, leukopenia, thrombocytopenia,heparin-induced thrombocytopenia (HIT), heparin-induced thrombocytopeniaand thrombosis (HITT), atrial fibrillation, implantation of a prostheticheart valve, genetic susceptibility to thrombosis, factor V Leiden,prothrombin gene mutation, methylenetetrahydrofolate reductase (MTHFR)polymorphism, platelet-receptor polymorphism, trauma, fractures, burns,or any combination thereof.

In an embodiment of the invention, may include administration of one ormore anti-coagulants or statins.

In an embodiment of the invention, the one or more anti-coagulants maybe selected from abciximab (ReoPro), acenocoumarol, antithrombin III,argatroban, aspirin, bivalirudin (Angiomax), clopidogrel, dabigatran,dabigatran etexilate (Pradaxa/Pradax), desirudin (Revasc/Iprivask),dipyridamole, eptifibatide (Integrilin), fondaparinux, heparin, hirudin,idraparinux, lepirudin (Refludan), low molecular weight heparin,melagatran, phenindione, phenprocoumon, ticlopidine, tirofiban(Aggrastat), warfarin, ximelagatran, ximelagatran (Exanta/Exarta), orany combination thereof.

In an embodiment of the invention, the one or more statins may beselected from pravastatin, lovastatin, simvastatin, fluvastatin,atorvastatin, simvastatin, nystatin, pentastatin, cerivastatin, or anycombination thereof.

In an embodiment of the invention, the patient's coagulation profile maybe assessed by measurement of the patient's serum level of one or moreof D-dimer, Factor II, Factor V, Factor VIII, Factor IX, Factor XI,Factor XII, F/fibrin degradation products, thrombin-antithrombin IIIcomplex, fibrinogen, plasminogen, prothrombin, and von Willebrandfactor.

In an embodiment of the invention, the patient's coagulation profile maybe assessed by a functional measurement of clotting ability.

In an embodiment of the invention, the functional measurement ofclotting ability may be selected from prothrombin time (PT), prothrombinratio (PR), international normalized ratio (INR), or any combinationthereof.

In an embodiment of the invention, the method may include measuring thepatient's international normalized ratio (INR) prior to administrationof the IL-6 antagonist, and administering to the patient an IL-6antagonist if the patient's INR is less than about 0.9.

In an embodiment of the invention, the invention may include measuringthe patient's international normalized ratio (INR) prior toadministration of the IL-6 antagonist, and administering to the patientan IL-6 antagonist if the patient's INR is less than about 0.5.

54.) In an embodiment of the invention, the patient's INR may be raisedto more than approximately 0.9 within 4 weeks of administering to thepatient an IL-6 antagonist.

In an embodiment of the invention, the method may include measuring thepatient's serum D-dimer level prior to administration of the IL-6antagonist, and administering the IL-6 antagonist if the patient's serumserum D-dimer level is above the normal reference range.

In an embodiment of the invention, the patient's serum D-dimer level maybe lowered to less than the upper limit of the normal reference rangewithin 4 weeks of administering to the patient an IL-6 antagonist.

In an embodiment of the invention, the method may result in a prolongedimprovement in the patient's coagulation profile.

In an embodiment of the invention, the patient's coagulation profile maybe measurably improved within about 2 weeks of administration of theIL-6 antagonist.

In an embodiment of the invention, the patient's coagulation profile mayremain measurably improved approximately 12 weeks after administering tothe patient an IL-6 antagonist.

In an embodiment of the invention, the patient's survivability may beimproved.

In an embodiment of the invention, the IL-6 antagonist may be anantisense nucleic acid.

In an embodiment of the invention, the IL-6 antagonist may be anantisense nucleic acid, for example comprising at least approximately 10nucleotides of a sequence encoding IL-6, IL-6 receptor alpha, gp130, p38MAP kinase, JAK1, JAK2, JAK3, or SYK.

In an embodiment of the invention, the antisense nucleic acid maycomprise DNA, RNA, peptide nucleic acid, locked nucleic acid, morpholino(phosphorodiamidate morpholino oligo), glycerol nucleic acid, threosenucleic acid, or any combination thereof.

In an embodiment of the invention, the IL-6 antagonist may compriseActemra (Tocilizumab), Remicade, Zenapax, or any combination thereof.

In an embodiment of the invention, the IL-6 antagonist may comprise apolypeptide having a sequence comprising a fragment of IL-6, IL-6receptor alpha, gp130, p38 MAP kinase, JAK1, JAK2, JAK3, SYK, or anycombination thereof, such as a fragment or full-length polypeptide thatis at least 40 amino acids in length.

In an embodiment of the invention, the IL-6 antagonist may comprise asoluble IL-6, IL-6 receptor alpha, gp130, p38 MAP kinase, JAK1, JAK2,JAK3, SYK, or any combination thereof.

In an embodiment of the invention, the IL-6 antagonist may be coupled toa half-life increasing moiety.

In an embodiment of the invention, the method may further compriseadministration of one or more statins to the patient, including withoutlimitation thereto pravastatin, lovastatin, simvastatin, fluvastatin,atorvastatin, simvastatin, nystatin, pentastatin, cerivastatin, or anycombination thereof.

Another embodiment of the invention relates to a composition comprisingan IL-6 antagonist, and an anti-coagulant. In an embodiment of theinvention, the one or more anti-coagulants may be selected fromabciximab (ReoPro), acenocoumarol, antithrombin III, argatroban,aspirin, bivalirudin (Angiomax), clopidogrel, dabigatran, dabigatranetexilate (Pradaxa/Pradax), desirudin (Revasc/Iprivask), dipyridamole,eptifibatide (Integrilin), fondaparinux, heparin, hirudin, idraparinux,lepirudin (Refludan), low molecular weight heparin, melagatran,phenindione, phenprocoumon, ticlopidine, tirofiban (Aggrastat),warfarin, ximelagatran, ximelagatran (Exanta/Exarta), or any combinationthereof.

Another embodiment of the invention relates to a composition comprisingan IL-6 antagonist, and a chemotherapy agent. In an embodiment of theinvention, the chemotherapy agent may be selected from VEGF antagonists,EGFR antagonists, platins, taxols, irinotecan, 5-fluorouracil,gemcytabine, leucovorine, steroids, cyclophosphamide, melphalan, vincaalkaloids (e.g., vinblastine, vincristine, vindesine and vinorelbine),mustines, tyrosine kinase inhibitors, radiotherapy, sex hormoneantagonists, selective androgen receptor modulators, selective estrogenreceptor modulators, PDGF antagonists, TNF antagonists, IL-1antagonists, interleukins (e.g. IL-12 or IL-2), IL-12R antagonists,Toxin conjugated monoclonal antibodies, tumor antigen specificmonoclonal antibodies, Erbitux, Avastin, Pertuzumab, anti-CD20antibodies, Rituxan, ocrelizumab, ofatumumab, DXL625, herceptin, or anycombination thereof.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

FIG. 1 shows that a variety of unique epitopes were recognized by thecollection of anti-IL-6 antibodies prepared by the antibody selectionprotocol. Epitope variability was confirmed by antibody-IL-6 bindingcompetition studies (ForteBio Octet).

FIG. 2 shows alignments of variable light and variable heavy sequencesbetween a rabbit antibody variable light and variable heavy sequencesand homologous human sequences and the final humanized sequences.Framework regions are identified FR1-FR4. Complementarity determiningregions are identified as CDR1-CDR3. Amino acid residues are numbered asshown. The initial rabbit sequences are called RbtVL and RbtVH for thevariable light and variable heavy sequences respectively. Three of themost similar human germline antibody sequences, spanning from Framework1 through to the end of Framework 3, are aligned below the rabbitsequences. The human sequence that is considered the most similar to therabbit sequence is shown first. In this example those most similarsequences are L12A for the light chain and 3-64-04 for the heavy chain.Human CDR3 sequences are not shown. The closest human Framework 4sequence is aligned below the rabbit Framework 4 sequence. The verticaldashes indicate a residue where the rabbit residue is identical with oneor more of the human residues at the same position. The bold residuesindicate that the human residue at that position is identical to therabbit residue at the same position. The final humanized sequences arecalled VLh and VHh for the variable light and variable heavy sequencesrespectively. The underlined residues indicate that the residue is thesame as the rabbit residue at that position but different than the humanresidues at that position in the three aligned human sequences.

FIG. 3 demonstrates the high correlation between the IgG produced andantigen specificity for an exemplary IL-6 protocol. 9 of 11 wells showedspecific IgG correlation with antigen recognition.

FIG. 4 provides the α-2-macroglobulin (A2M) dose response curve forantibody Ab1 administered intravenously at different doses one hourafter a 100 μg/kg s.c. dose of human IL-6.

FIG. 5 provides survival data for the antibody Ab1 progression groupsversus control groups.

FIG. 6 provides additional survival data for the antibody Ab1 regressiongroups versus control groups.

FIG. 7 provides survival data for polyclonal human IgG at 10 mg/kg i.v.every three days (270-320 mg tumor size) versus antibody Ab1 at 10 mg/kgi.v. every three days (270-320 mg tumor size).

FIG. 8 provides survival data for polyclonal human IgG at 10 mg/kg i.v.every three days (400-527 mg tumor size) versus antibody Ab1 at 10 mg/kgi.v. every three days (400-527 mg tumor size).

FIG. 9 provides a pharamcokinetic profile of antibody Ab1 in cynomolgusmonkey. Plasma levels of antibody Ab1 were quantitated through antigencapture ELISA. This protein displays a half life of between 12 and 17days consistent with other full length humanized antibodies.

FIG. 10 (A-D) provides binding data for antibodies Ab4, Ab3, Ab8 andAb2, respectively. FIG. 10 E provides binding data for antibodies Ab1,Ab6 and Ab7.

FIG. 11 summarizes the binding data of FIG. 10 (A-E) in tabular form.

FIG. 12 presents the sequences of the 15 amino acid peptides used in thepeptide mapping experiment of Example 14.

FIG. 13 presents the results of the blots prepared in Example 14.

FIG. 14 presents the results of the blots prepared in Example 14.

FIG. 15A shows affinity and binding kinetics of Ab1 for IL-6 of variousspecies.

FIG. 15B demonstrates inhibition of IL-6 by Ab1 in the T1165 cellproliferation assay.

FIG. 16. shows the mean plasma concentration of Ab1 resulting from asingle administration of Ab1 to healthy male subjects in several dosagegroups.

FIG. 17 shows mean area under the plasma Ab1 concentration time curve(AUC) for the dosage groups shown in FIG. 16.

FIG. 18 shows mean peak plasma Ab1 concentration (C_(max)) for thedosage groups shown in FIG. 16.

FIG. 19 summarizes Ab1 pharmacokinetic measurements of the dosage groupsshown in FIG. 16.

FIG. 20 shows the mean plasma concentration of Ab1 resulting from asingle administration of Ab1 to patients with advanced cancer.

FIG. 21 illustrates the unprecedented elimination half-life of Ab1compared with other anti-IL-6 antibodies.

FIG. 22 shows increased hemoglobin concentration followingadministration of Ab1 to patients with advanced cancer.

FIG. 23 shows mean plasma lipid concentrations following administrationof Ab1 to patients with advanced cancer.

FIG. 24 shows mean neutrophil counts following administration of Ab1 topatients with advanced cancer.

FIG. 25 demonstrates suppression of serum CRP levels in healthyindividuals.

FIG. 26 (A-B) demonstrates suppression of serum CRP levels in advancedcancer patients.

FIG. 27 shows prevention of weight loss by Ab1 in a mouse cancercachexia model.

FIG. 28 shows the physical appearance of representative Ab1-treated andcontrol mice in a cancer cachexia model.

FIG. 29 demonstrates that Ab1 promotes weight gain in advanced cancerpatients.

FIG. 30 demonstrates that Ab1 reduces fagitue in advanced cancerpatients.

FIG. 31 demonstrates that Ab1 promotes hand grip strength in advancedcancer patients.

FIG. 32 demonstrates that Ab1 suppresses an acute phase protein (SerumAmyloid A) in mice.

FIG. 33 demonstrates that Ab1 increase plasma albumin concentration inadvanced cancer patients.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS Definitions

It is to be understood that this invention is not limited to theparticular methodology, protocols, cell lines, animal species or genera,and reagents described, as such may vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to limit the scope ofthe present invention which will be limited only by the appended claims.

As used herein the singular forms “a”, “and”, and “the” include pluralreferents unless the context clearly dictates otherwise. Thus, forexample, reference to “a cell” includes a plurality of such cells andreference to “the protein” includes reference to one or more proteinsand equivalents thereof known to those skilled in the art, and so forth.All technical and scientific terms used herein have the same meaning ascommonly understood to one of ordinary skill in the art to which thisinvention belongs unless clearly indicated otherwise.

Interleukin-6 (IL-6): As used herein, interleukin-6 (IL-6) encompassesnot only the following 212 amino acid sequence available as GenBankProtein Accession No. NP_000591:

(SEQ ID NO: 1) MNSFSTSAFGPVAFSLGLLLVLPAAFPAPVPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWLQDMTTHLILRSFKE FLQSSLRALRQMbut also any pre-pro, pro- and mature forms of this IL-6 amino acidsequence, as well as mutants and variants including allelic variants ofthis sequence.

IL-6 Antagonist: As used herein, the terms “IL-6 antagonist,” andgrammatical variants thereof include any composition that prevents,inhibits, or lessens the effect(s) of IL-6 signaling. Generally, suchantagonists may reduce the levels or activity of IL-6, IL-6 receptoralpha, gp130, or a molecule involved in IL-6 signal transduction, or mayreduce the levels or activity complexes between the foregoing (e.g.,reducing the activity of an IL-6/IL-6 receptor complex). Antagonistsinclude antisense nucleic acids, including DNA, RNA, or a nucleic acidanalogue such as a peptide nucleic acid, locked nucleic acid, morpholino(phosphorodiamidate morpholino oligo), glycerol nucleic acid, or threosenucleic acid. See Heasman, Dev Biol. 2002 Mar. 15; 243(2):209-14; Hannonand Rossi, Nature. 2004 Sep. 16; 431(7006):371-8; Paul et al., NatBiotechnol. 2002 May; 20(5):505-8; Zhang et al., J Am Chem Soc. 2005Mar. 30; 127(12):4174-5; Wahlestedt et al., Proc Natl Acad Sci USA. 2000May 9; 97(10):5633-8; Hanvey et al., 1992 Nov. 27; 258(5087):1481-5;Braasch et al., Biochemistry. 2002 Apr. 9; 41(14):4503-10; Schoning etal., Science. 2000 Nov. 17; 290(5495):1347-51. In addition IL-6antagonists specifically include peptides that block IL-6 signaling suchas those described in any of U.S. Pat. Nos. 6,599,875; 6,172,042;6,838,433; 6,841,533; 5,210,075 et al. Also, IL-6 antagonists accordingto the invention may include p38 MAP kinase inhibitors such as thosereported in US20070010529 et al. given this kinase's role in cytokineproduction and more particularly IL-6 production. Further, IL-6antagonosts according to the invention include the glycoalkaloidcompounds reported in US20050090453 as well as other IL-6 antagonistcompounds isolatable using the IL-6 antagononist screening assaysreported therein. Other IL-6 antagonists include antibodies, such asanti-IL-6 antibodies, anti-IL-6 receptor alpha antibodies, anti-gp130antibodies, and anti-p38 MAP kinase antibodies including (but notlimited to) the anti-IL-6 antibodies disclosed herein, Actemra(Tocilizumab), Remicade, Zenapax, or any combination thereof. Other IL-6antagonists include portions or fragments of molecules involved in IL-6signaling, such as IL-6, IL-6 receptor alpha, and gp130, which may benative, mutant, or variant sequence, and may optionally be coupled toother moieties (such as half-life-increasing moieties, e.g. an Fcdomain). For example, an IL-6 antagonist may be a soluble IL-6 receptoror fragment, a soluble IL-6 receptor:Fc fusion protein, a small moleculeinhibitor of IL-6, an anti-IL-6 receptor antibody or antibody fragment,antisense nucleic acid, etc. Other IL-6 antagonists include avemirs,such as C326 (Silverman et al., Nat Biotechnol. 2005 December;23(12):1556-61) and small molecules, such as synthetic retinoid AM80(tamibarotene) (Takeda et al., Arterioscler Thromb Vasc Biol. 2006 May;26(5):1177-83). Such IL-6 antagonists may be administered by any meansknown in the art, including contacting a subject with nucleic acidswhich encode or cause to be expressed any of the foregoing polypeptidesor antisense sequences.

Thrombosis: As used herein, thrombosis refers to a thrombus (blood clot)inside a blood vessel. The term encompasses, without limitation,arterial and venous thrombosis, including deep vein thrombosis, portalvein thrombosis, jugular vein thrombosis, renal vein thrombosis, stroke,myocardial infarction, Budd-Chiari syndrome, Paget-Schroetter disease,and cerebral venous sinus thrombosis. Diseases and conditions associatedwith thrombosis include, without limitation, acute venous thrombosis,pulmonary embolism, thrombosis during pregnancy, hemorrhagic skinnecrosis, acute or chronic disseminated intravascular coagulation (DIC),clot formation from surgery, long bed rest, long periods ofimmobilization, venous thrombosis, fulminant meningococcemia, acutethrombotic stroke, acute coronary occlusion, acute peripheral arterialocclusion, massive pulmonary embolism, axillary vein thrombosis, massiveiliofemoral vein thrombosis, occluded arterial cannulae, occluded venouscannulae, cardiomyopathy, venoocclusive disease of the liver,hypotension, decreased cardiac output, decreased vascular resistance,pulmonary hypertension, diminished lung compliance, leukopenia, andthrombocytopenia.

D-Dimer: As used herein, D-dimer refers to a fibrin degradation productproduced during the break down of blood clots by the enzyme plasmin.Monoclonal antibodies specifically reactive against D-dimer are readilyavailable, e.g. DD-3B6/22 (Elms et al., 1986, Am J Clin Pathol.85:360-4). Clinical measurements of D-dimer levels are routinelyperformed, e.g., using a red blood cell agglutination test, ELISA, etc.(reviewed in Dempfle, Semin Vasc Med, 2005 November; 5(4):315-20).Measurements of D-dimer may vary depending on the measurement method andtesting lab; nonetheless, a normal “reference range” may be readilyestablished for any particular method and testing lab, e.g. by takingmeasurements from healthy individuals. Accordingly, an elevated D-dimerlevel is understood by persons skilled in the art to refer to a D-dimerlevel that is above the reference range for the partcular method andtesting lab.

Coagulation profile: As used herein, coagulation profile refersgenerally to the functioning of the coagulation system. Both the tissuefactor (extrinsic) and contact activation (intrinsic) pathways ofclotting are components of the coagulation profile. A normal coagulationprofile refers to coagulation functioning as in a normal, healthyindividual, i.e., maintaining balance between ability to controlbleeding and tendency towards excessive clotting (thrombotic tendency).An abnormal coagulation profile may be a decrease or an increase incoagulation tendency. One particularly abnormal coagulation profile ishypercoagulation, which refers to a greatly increased risk of excessiveclot formation, resulting in high risk of thrombosis. Coagulationprofile may be assessed by various tests and assays known in the art,such as: the activated partial thromboplastin time (aPTT) test;prothrombin time (PT) test (typical reference range of 12 to 15 second);measurements derived from the PT test, such as prothrombin ratio (PR)and international normalized ratio (INR) (typical reference range 0.8 to1.2); fibrinogen testing (e.g. the Clauss method (Clauss A, “RapidPhysiological Coagulation Method for the Determination of Fibrinogen[German],” Acta Haematol, 1957, 17:237-46) or the Ellis method (Ellis BC and Stransky A, “A Quick and Accurate Method for the Determination ofFibrinogen in Plasma,” J Lab Clin Med, 1961, 58:477-88); assays foractivated protein C resistance, protein C, protein S, and antithrombin;assays for antiphospholipid antibodies (lupus anticoagulant andanticardiolipin antibodies); elevated homocysteine; assays forplasminogen, dysfibrinogenemia, heparin cofactor II, or platelethyperaggregability. Other assays useful to assess coagulation profileinclude measurement of clotting factors and/or indicators of clotting,such as serum levels of D-dimer, Factor II, Factor V, Factor VIII,Factor IX, Factor XI, Factor XII, F/fibrin degradation products,thrombin-antithrombin III complex, thrombocytosis, fibrinogen,plasminogen, prothrombin, and von Willebrand factor. Worsening incoagulation profile refers to a measureable change in an indicator ofcoagulation, e.g., any of the aforementioned assays, that reflects adeterioration of the normal coagulation tendency, such that the measuredvalue becomes abnormal or deviates farther from the normal range thanpreviously. Improvement in coagulation profile refers to a measureablechange in an indicator of coagulation, e.g., any of the aforementionedassays, that reflects a partial or full restoration of the normalcoagulation tendency, i.e., after a therapeutic intervention, such asadministration of an anti-IL-6 antibody, the measured value is in thenormal range or closer to the normal range than prior to the therapeuticintervention.

Disease or condition: As used herein, “disease or condition” refers to adisease or condition that a patient has been diagnosed with or issuspected of having, particularly a disease or condition associated withelevated IL-6. A disease or condition encompasses, without limitationthereto, the side-effects of medications or treatments (such asradiation therapy), as well as idiopathic conditions characterized bysymptoms that include elevated IL-6.

Cachexia: As used herein, cachexia, also known as wasting disease,refers to any disease marked especially by progressive emaciation,weakness, general ill health, malnutrition, loss of body mass, loss ofmuscle mass, or an accelerated loss of skeletal muscle in the context ofa chronic inflammatory response (reviewed in Kotler, Ann Intern Med.2000 Oct. 17; 133(8):622-34). Diseases and conditions in which cachexiais frequently observed include cancer, rheumatoid arthritis, AIDS, heartdisease, dehydration, malnutrition, lead exposure, malaria, respiratorydisease, old age, hypothyroidism, tuberculosis, hypopituitarism,neurasthenia, hypernatremia, hyponatremia, renal disease, splenica,ankylosing spondylitis, failure to thrive (faltering growth) and otherdiseases, particularly chronic diseases. Cachexia may also be idiopathic(arising from an uncertain cause). Weight assessment in a patient isunderstood to exclude growths or fluid accumulations, e.g. tumor weight,extravascular fluid accumulation, etc. Cachexia may be assessed bymeasureent of a patient's total body mass (exclusive of growths or fluidaccumulations), total lean (fat-free) body mass, lean mass of the armsand legs (appendicular lean mass, e.g. measured using dual-energy x-rayabsorptiometry or bioelectric impedance spectroscopy), and/or lean bodymass index (lean body mass divided by the square of the patient'sheight). See Kotler, Ann Intern Med. 2000 Oct. 17; 133(8):622-34;Marcora et al., Rheumatology (Oxford). 2006 November; 45(11):1385-8.

Weakness: As used herein, weakness refers physical fatigue, whichtypically manifests as a loss of muscle strength and/or endurance.Weakness may be central (affecting most or all of the muscles in thebody) or peripheral (affecting a subset of muscles). Weakness includes“true weakness,” in which a patient's muscles have a decrease in somemeasure of peak and/or sustained force output, and “perceived weakness,”in which a patient perceives that a greater effort is required forperformance of a task even though objectively measured strength remainsnearly the same, and may be objectively measured or self-reported by thepatient. For example, weakness may be objectively measured using thehand grip strength test (a medically recognized test for evaluatingmuscle strength), typically employing a handgrip dynamometer.

Fatigue: As used herein, fatigue refers to mental fatigue (for physicalfatigue see “weakness”). Fatigue includes drowsiness (somnolence) and/ordecreased attention. Fatigue may be measured using a variety of testsknown in the art, such as the FACIT-F (Functional Assessment of ChronicIllness Therapy-Fatigue) test. See, e.g., Cella, D., Lai, J. S., Chang,C. H., Peterman, A., & Slavin, M. (2002). Fatigue in cancer patientscompared with fatigue in the general population. Cancer, 94(2), 528-538;Cella, D., Eton, D. T., Lai, F J-S., Peterman, A. H & Merkel, D. E.(2002). Combining anchor and distribution based methods to deriveminimal clinically important differences on the Functional Assessment ofCancer Therapy anemia and fatigue scales. Journal of Pain & SymptomManagement, 24 (6) 547-561.

Fever: As used herein, “fever” refers to a body temperature set-pointthat is elevated by at least 1 to 2 degrees Celsius. Fever is oftenassociated with a subjective feeling of hypothermia exhibited as a coldsensation, shivering, increased heart rate and respiration rate by whichthe individual's body reaches the increased set-point. As is wellunderstood in the medical arts, normal body temperature typically varieswith activity level and time of day, with highest temperatures observedin the afternoon and early evening hours, and lowest temperaturesobserved during the second half of the sleep cycle, and temperaturemeasurements may be influenced by external factors such as mouthbreathing, consumption of food or beverage, smoking, or ambienttemperature (depending on the type of measurement). Moreover, the normaltemperature set point for individuals may vary by up to about 0.5degrees Celsius, thus a medical professional may interpret anindividual's temperature in view of these factors to diagnose whether afever is present. Generally speaking, a fever is typically diagnosed bya core body temperature above 38.0 degrees Celsius, an oral temperatureabove 37.5 degrees Celsius, or an axillary temperature above 37.2degrees Celsius.

Improved: As used herein, “improved,” “improvement,” and othergrammatical variants, includes any beneficial change resulting from atreatment. A beneficial change is any way in which a patient's conditionis better than it would have been in the absence of the treatment.“Improved” includes prevention of an undesired condition, slowing therate at which a condition worsens, delaying the development of anundesired condition, and restoration to an essentially normal condition.For example, improvement in cachexia encompasses any increase inpatient's mass, such as total body mass (excluding weight normallyexcluded during assessment of cachexia, e.g. tumor weight, extravascularfluid accumulation, etc.), lean body mass, and/or appendicular leanmass, as well as any delay or slowing in the rate of loss of mass, orprevention or slowing of loss of mass associated with a disease orcondition with which the patient has been diagnosed. For anotherexample, improvement in weakness encompasses any increase in patient'sstrength, as well as any delay or slowing in the rate of loss ofstrength, or prevention or slowing of loss of strength associated with adisease or condition with which the patient has been diagnosed. For yetanother example, improvement in fatigue encompasses any decrease inpatient's fatigue, as well as any delay or slowing in the rate ofincrease of fatigue, or prevention or slowing of increase in fatigueassociated with a disease or condition with which the patient has beendiagnosed. For still another example, improvement in fever encompassesany decrease in patient's fever, as well as any delay or slowing in therate of increase in fever, or prevention or slowing of increase in feverassociated with a disease or condition with which the patient has beendiagnosed.

C-Reactive Protein (CRP): As used herein, C-Reactive Protein (CRP)encompasses not only the following 224 amino acid sequence available asGenBank Protein Accession No. NP_000558:MEKLLCFLVLTSLSHAFGQTDMSRKAFVFPKESDTSYVSLKAPLTKPLKAFTVCLHFYTELSSTRGYSIFSYATKRQDNEILIFWSKDIGYSFTVGGSEILFEVPEVTVAPVHICTSWESASGIVEFWVDGKPRVRKSLKKGYTVGAEASIILGQEQDSFGGNFEGSQSLVGDIGNVNMWDFVLSPDEINTIYLGGPFSPNVLNWRALKYEVQGEVFTKPQ LWP (SEQ ID NO:647), but also any pre-pro, pro- and mature forms of this CRP amino acidsequence, as well as mutants and variants including allelic variants ofthis sequence. CRP levels, e.g. in the serum, liver, tumor, or elsewherein the body, can be readily measured using routine methods andcommercially available reagents, e.g. ELISA, antibody test strip,immunoturbidimetry, rapid immunodiffusion, visual agglutination, Westernblot, Northern blot, etc. As mentioned above CRP levels may in additionbe measured in patients having or at risk of developing thrombosisaccording to the invention.

Interleukin-6 receptor (IL-6R); also called IL-6 receptor alpha(IL-6RA): As used herein, “interleukin-6 receptor” (“IL-6R”; also “IL-6receptor alpha” or “IL-6RA”) encompasses not only the following 468amino acid sequence available as Swiss-Prot Protein Accession No.P08887: MLAVGCALLAALLAAPGAALAPRRCPAQEVARGVLTSLPGDSVTLTCPGVEPEDNATVHWVLRKPAAGSHPSRWAGMGRRLLLRSVQLHDSGNYSCYRAGRPAGTVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSLTTKAVLLVRKFQNSPAEDFQEPCQYSQESQKFSCQLAVPEGDSSFYIVSMCVASSVGSKFSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTWQDPHSWNSSFYRLRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQGEWSEWSPEAMGTPWTESRSPPAENEVSTPMQALTTNKDDDNILFRDSANATSLPVQDSSSVPLPTFLVAGGSLAFGTLLCIAIVLRFKKTWKLRALKEGKTSMHPPYSLGQLVPERPRPTPVLVPLISPPVSPSSLGSDNTSSHNRPDARDPRSPYDISNTDYFFPR (SEQ ID NO: 648), but also anypre-pro, pro- and mature forms of this amino acid sequence, as well asmutants and variants including allelic variants of this sequence.

gp130: As used herein, gp130 (also called Interleukin-6 receptor subunitbeta) encompasses not only the following 918 precursor amino acidsequence available as Swiss-Prot Protein Accession No. P40189:MLTLQTWVVQALFIFLTTESTGELLDPCGYISPESPVVQLHSNFTAVCVLKEKCMDYFHVNANYIVWKTNHFTIPKEQYTIINRTASSVTFTDIASLNIQLTCNILTFGQLEQNVYGITIISGLPPEKPKNLSCIVNEGKKMRCEWDGGRETHLETNFTLKSEWATHKFADCKAKRDTPTSCTVDYSTVYFVNIEVWVEAENALGKVTSDHINFDPVYKVKPNPPHNLSVINSEELSSILKLTWTNPSIKSVIILKYNIQYRTKDASTWSQIPPEDTASTRSSFTVQDLKPFTEYVFRIRCMKEDGKGYWSDWSEEASGITYEDRPSKAPSFWYKIDPSHTQGYRTVQLVWKTLPPFEANGKILDYEVTLTRWKSHLQNYTVNATKLTVNLTNDRYLATLTVRNLVGKSDAAVLTIPACDFQATHPVMDLKAFPKDNMLWVEWTTPRESVKKYILEWCVLSDKAPCITDWQQEDGTVHRTYLRGNLAESKCYLITVTPVYADGPGSPESIKAYLKQAPPSKGPTVRTKKVGKNEAVLEWDQLPVDVQNGFIRNYTIFYRTIIGNETAVNVDSSHTEYTLSSLTSDTLYMVRMAAYTDEGGKDGPEFTFTTPKFAQGEIEAIVVPVCLAFLLTTLLGVLFCFNKRDLIKKHIWPNVPDP SKSHIAQWSPHTPPRHNFNSKDQMYSDGNFTDVSVVEIEANDKKPFPEDLKSLDLFKKEKINTEGHSSGIGGSSCMSSSRPSISSSDENESSQNTSSTVQYSTVVHSGYRHQVPSVQVFSRSESTQPLLDSEERPEDLQLVDHVDGGDGILPRQQYFKQNCSQHESSPDISHFERSKQVSSVNEEDFVRLKQQISDHISQSCGSGQMKMFQEVSAADAFGPGTEGQVERFETVGMEAATDEGMPKSYLPQTVRQGGYMPQ (SEQ ID NO: 649), but also anypre-pro, pro- and mature forms of this amino acid sequence, such as themature form encoded by amino acids 23 through 918 of the sequence shown,as well as mutants and variants including allelic variants of thissequence.

Glasgow Prognostic Score (GPS): As used herein, Glasgow Prognostic Score(GPS) refers to an inflammation-based prognostic score that awards onepoint for a serum albumin level less than <35 mg/L and one point for aCRP level above 10 mg/L. Thus, a GPS of 0 indicates normal albumin andCRP, a GPS of 1 indicates reduced albumin or elevated CRP, and a GPS of2 indicates both reduced albumin and elevated CRP.

Effective amount: As used herein, “effective amount,” “amount effectiveto,” “amount of X effective to” and the like, refer to an amount of anactive ingredient that is effective to relieve or reduce to some extentone or more of the symptoms of the disease in need of treatment, or toretard initiation of clinical markers or symptoms of a disease in needof prevention, when the compound is administered. Thus, an effectiveamount refers to an amount of the active ingredient which exhibiteffects such as (i) reversing the rate of progress of a disease; (ii)inhibiting to some extent further progress of the disease; and/or, (iii)relieving to some extent (or, preferably, eliminating) one or moresymptoms associated with the disease. The effective amount may beempirically determined by experimenting with the compounds concerned inknown in vivo and in vitro model systems for a disease in need oftreatment. The context in which the phrase “effective amount” is usedmay indicate a particular desired effect. For example, “an amount of ananti-IL-6 antibody effective to prevent or treat a hypercoagulablestate” and similar phrases refer to an amount of anti-IL-6 antibodythat, when administered to a subject, will cause a measurableimprovement in the subject's coagulation profile, or prevent, slow,delay, or arrest, a worsening of the coagulation profile for which thesubject is at risk. An effective amount will vary according to theweight, sex, age and medical history of the individual, as well as theseverity of the patient's condition(s), the type of disease(s), mode ofadministration, and the like. An effective amount may be readilydetermined using routine experimentation, e.g., by titration(administration of increasing dosages until an effective dosage isfound) and/or by reference to amounts that were effective for priorpatients. Generally, the anti-IL-6 antibodies of the present inventionwill be administered in dosages ranging between about 0.1 mg/kg andabout 20 mg/kg of the patient's body-weight.

Prolonged improvement in coagulation profile: As used herein, “prolongedimprovement in coagulation profile” and similar phrases refer to ameasurable improvement in the subject's coagulation profile relative tothe initial coagulation profile (i.e. the coagulation profile at a timebefore treatment begins) that is detectable within about a week fromwhen treatment begins (e.g. administration of an IL-6 antagonist) andremains improved for a prolonged duration, e.g., at least about 14 days,at least about 21 days, at least about 28 days, at least about 35 days,at least about 40 days, at least about 50 days, at least about 60 days,at least about 70 days, at least about 11 weeks, or at least about 12weeks from when the treatment begins.

Prolonged reduction in serum CRP: As used herein, “prolonged reductionin serum CRP” and similar phrases refer to a measurable decrease inserum CRP level relative to the initial serum CRP level (i.e. the serumCRP level at a time before treatment begins) that is detectable withinabout a week from when a treatment begins (e.g. administration of ananti-IL-6 antibody) and remains below the initial serum CRP level for anprolonged duration, e.g. at least about 14 days, at least about 21 days,at least about 28 days, at least about 35 days, at least about 40 days,at least about 50 days, at least about 60 days, at least about 70 days,at least about 11 weeks, or at least about 12 weeks from when thetreatment begins.

Prolonged increase in serum albumin: As used herein, “prolonged increasein serum albumin” and similar phrases refer to a measurable decrease inserum albumin level relative to the initial serum albumin level (i.e.the serum albumin level at a time before treatment begins) that isdetectable within about a week from when a treatment begins (e.g.administration of an anti-IL-6 antibody) and remains above the initialserum albumin level for an prolonged duration, e.g. at least about 14days, at least about 21 days, at least about 28 days, at least about 35days, at least about 40 days, at least about 50 days, at least about 60days, at least about 70 days, at least about 11 weeks, or at least about12 weeks from when the treatment begins.

Prolonged improvement in cachexia: As used herein, “prolongedimprovement in cachexia” refers to a measureable improvement patient'sbody mass, lean body mass, apendicular lean body mass, and/or lean bodymass index, relative to the initial level (i.e. the level at a timebefore treatment begins) that is detectable within about 4 weeks andremains improved for a prolonged duration, e.g. at least about 35 days,at least about 40 days, at least about 50 days, at least about 60 days,at least about 70 days, at least about 11 weeks, or at least about 12weeks from when the treatment begins.

Prolonged improvement in weakness: As used herein, “prolongedimprovement in weakness” refers to a measureable improvement in muscularstrength, relative to the initial level (i.e. the level at a time beforetreatment begins) that is detectable within about 2 weeks and remainsimproved for a prolonged duration, e.g. at least about 21 days, at leastabout 28 days, at least about 35 days, at least about 40 days, at leastabout 50 days, at least about 60 days, at least about 70 days, at leastabout 11 weeks, or at least about 12 weeks from when the treatmentbegins.

Prolonged improvement in fatigue: As used herein, “prolonged improvementin fatigue” refers to a measureable improvement in fatigue, relative tothe initial level (i.e. the level at a time before treatment begins)that is detectable within about 1 week and remains improved for aprolonged duration, e.g. at least about 14 days, at least about 21 days,at least about 28 days, at least about 35 days, at least about 40 days,at least about 50 days, at least about 60 days, at least about 70 days,at least about 11 weeks, or at least about 12 weeks from when thetreatment begins.

Prolonged improvement in fever: As used herein, “prolonged improvementin fever” refers to a measureable decrease in fever (e.g. peaktemperature or amount of time that temperature is elevated), relative tothe initial level (i.e. the level at a time before treatment begins)that is detectable within about 1 week and remains improved for aprolonged duration, e.g. at least about 14 days, at least about 21 days,at least about 28 days, at least about 35 days, at least about 40 days,at least about 50 days, at least about 60 days, at least about 70 days,at least about 11 weeks, or at least about 12 weeks from when thetreatment begins.

Mating competent yeast species: In the present invention this isintended to broadly encompass any diploid or tetraploid yeast which canbe grown in culture. Such species of yeast may exist in a haploid,diploid, or tetraploid form. The cells of a given ploidy may, underappropriate conditions, proliferate for indefinite number of generationsin that form. Diploid cells can also sporulate to form haploid cells.Sequential mating can result in tetraploid strains through furthermating or fusion of diploid strains. In the present invention thediploid or polyploidal yeast cells are preferably produced by mating orspheroplast fusion.

In one embodiment of the invention, the mating competent yeast is amember of the Saccharomycetaceae family, which includes the generaArxiozyma; Ascobotryozyma; Citeromyces; Debaryomyces; Dekkera;Eremothecium; Issatchenkia; Kazachstania; Kluyveromyces; Kodamaea;Lodderomyces; Pachysolen; Pichia; Saccharomyces; Saturnispora;Tetrapisispora; Torulaspora; Williopsis; and Zygosaccharomyces. Othertypes of yeast potentially useful in the invention include Yarrowia,Rhodosporidium, Candida, Hansenula, Filobasium, Filobasidellla,Sporidiobolus, Bullera, Leucosporidium and Filobasidella.

In a preferred embodiment of the invention, the mating competent yeastis a member of the genus Pichia. In a further preferred embodiment ofthe invention, the mating competent yeast of the genus Pichia is one ofthe following species: Pichia pastoris, Pichia methanolica, andHansenula polymorpha (Pichia angusta). In a particularly preferredembodiment of the invention, the mating competent yeast of the genusPichia is the species Pichia pastoris.

Haploid Yeast Cell: A cell having a single copy of each gene of itsnormal genomic (chromosomal) complement.

Polyploid Yeast Cell: A cell having more than one copy of its normalgenomic (chromosomal) complement.

Diploid Yeast Cell: A cell having two copies (alleles) of essentiallyevery gene of its normal genomic complement, typically formed by theprocess of fusion (mating) of two haploid cells.

Tetraploid Yeast Cell: A cell having four copies (alleles) ofessentially every gene of its normal genomic complement, typicallyformed by the process of fusion (mating) of two haploid cells.Tetraploids may carry two, three, four, or more different expressioncassettes. Such tetraploids might be obtained in S. cerevisiae byselective mating homozygotic heterothallic a/a and alpha/alpha diploidsand in Pichia by sequential mating of haploids to obtain auxotrophicdiploids. For example, a [met his] haploid can be mated with [ade his]haploid to obtain diploid [his]; and a [met arg] haploid can be matedwith [ade arg] haploid to obtain diploid [arg]; then the diploid[his]×diploid [arg] to obtain a tetraploid prototroph. It will beunderstood by those of skill in the art that reference to the benefitsand uses of diploid cells may also apply to tetraploid cells.

Yeast Mating: The process by which two haploid yeast cells naturallyfuse to form one diploid yeast cell.

Meiosis: The process by which a diploid yeast cell undergoes reductivedivision to form four haploid spore products. Each spore may thengerminate and form a haploid vegetatively growing cell line.

Selectable Marker: A selectable marker is a gene or gene fragment thatconfers a growth phenotype (physical growth characteristic) on a cellreceiving that gene as, for example through a transformation event. Theselectable marker allows that cell to survive and grow in a selectivegrowth medium under conditions in which cells that do not receive thatselectable marker gene cannot grow. Selectable marker genes generallyfall into several types, including positive selectable marker genes suchas a gene that confers on a cell resistance to an antibiotic or otherdrug, temperature when two ts mutants are crossed or a ts mutant istransformed; negative selectable marker genes such as a biosyntheticgene that confers on a cell the ability to grow in a medium without aspecific nutrient needed by all cells that do not have that biosyntheticgene, or a mutagenized biosynthetic gene that confers on a cellinability to grow by cells that do not have the wild type gene; and thelike. Suitable markers include but are not limited to: ZEO; G418; LYS3;MET1; MET3a; ADE1; ADE3; URA3; and the like.

Expression Vector: These DNA vectors contain elements that facilitatemanipulation for the expression of a foreign protein within the targethost cell. Conveniently, manipulation of sequences and production of DNAfor transformation is first performed in a bacterial host, e.g. E. coli,and usually vectors will include sequences to facilitate suchmanipulations, including a bacterial origin of replication andappropriate bacterial selection marker. Selection markers encodeproteins necessary for the survival or growth of transformed host cellsgrown in a selective culture medium. Host cells not transformed with thevector containing the selection gene will not survive in the culturemedium. Typical selection genes encode proteins that (a) conferresistance to antibiotics or other toxins, (b) complement auxotrophicdeficiencies, or (c) supply critical nutrients not available fromcomplex media. Exemplary vectors and methods for transformation of yeastare described, for example, in Burke, D., Dawson, D., & Stearns, T.(2000). Methods in yeast genetics: a Cold Spring Harbor Laboratorycourse manual. Plainview, N.Y.: Cold Spring Harbor Laboratory Press.

Expression vectors for use in the methods of the invention will furtherinclude yeast specific sequences, including a selectable auxotrophic ordrug marker for identifying transformed yeast strains. A drug marker mayfurther be used to amplify copy number of the vector in a yeast hostcell.

The polypeptide coding sequence of interest is operably linked totranscriptional and translational regulatory sequences that provide forexpression of the polypeptide in yeast cells. These vector componentsmay include, but are not limited to, one or more of the following: anenhancer element, a promoter, and a transcription termination sequence.Sequences for the secretion of the polypeptide may also be included,e.g. a signal sequence, and the like. A yeast origin of replication isoptional, as expression vectors are often integrated into the yeastgenome.

In one embodiment of the invention, the polypeptide of interest isoperably linked, or fused, to sequences providing for optimizedsecretion of the polypeptide from yeast diploid cells.

Nucleic acids are “operably linked” when placed into a functionalrelationship with another nucleic acid sequence. For example, DNA for asignal sequence is operably linked to DNA for a polypeptide if it isexpressed as a preprotein that participates in the secretion of thepolypeptide; a promoter or enhancer is operably linked to a codingsequence if it affects the transcription of the sequence. Generally,“operably linked” means that the DNA sequences being linked arecontiguous, and, in the case of a secretory leader, contiguous and inreading frame. However, enhancers do not have to be contiguous. Linkingis accomplished by ligation at convenient restriction sites oralternatively via a PCR/recombination method familiar to those skilledin the art (Gateway® Technology; Invitrogen, Carlsbad Calif.). If suchsites do not exist, the synthetic oligonucleotide adapters or linkersare used in accordance with conventional practice.

Promoters are untranslated sequences located upstream (5′) to the startcodon of a structural gene (generally within about 100 to 1000 bp) thatcontrol the transcription and translation of particular nucleic acidsequences to which they are operably linked. Such promoters fall intoseveral classes: inducible, constitutive, and repressible promoters(that increase levels of transcription in response to absence of arepressor). Inducible promoters may initiate increased levels oftranscription from DNA under their control in response to some change inculture conditions, e.g., the presence or absence of a nutrient or achange in temperature.

The yeast promoter fragment may also serve as the site for homologousrecombination and integration of the expression vector into the samesite in the yeast genome; alternatively a selectable marker is used asthe site for homologous recombination. Pichia transformation isdescribed in Cregg et al. (1985) Mol. Cell. Biol. 5:3376-3385.

Examples of suitable promoters from Pichia include the AOX1 and promoter(Cregg et al. (1989) Mol. Cell. Biol. 9:1316-1323); ICL1 promoter(Menendez et al. (2003) Yeast 20(13):1097-108);glyceraldehyde-3-phosphate dehydrogenase promoter (GAP) (Waterham et al.(1997) Gene 186(1):37-44); and FLD1 promoter (Shen et al. (1998) Gene216(1):93-102). The GAP promoter is a strong constitutive promoter andthe AOX and FLD1 promoters are inducible.

Other yeast promoters include ADH1, alcohol dehydrogenase II, GAL4,PHO3, PHO5, Pyk, and chimeric promoters derived therefrom. Additionally,non-yeast promoters may be used in the invention such as mammalian,insect, plant, reptile, amphibian, viral, and avian promoters. Mosttypically the promoter will comprise a mammalian promoter (potentiallyendogenous to the expressed genes) or will comprise a yeast or viralpromoter that provides for efficient transcription in yeast systems.

The polypeptides of interest may be produced recombinantly not onlydirectly, but also as a fusion polypeptide with a heterologouspolypeptide, e.g. a signal sequence or other polypeptide having aspecific cleavage site at the N-terminus of the mature protein orpolypeptide. In general, the signal sequence may be a component of thevector, or it may be a part of the polypeptide coding sequence that isinserted into the vector. The heterologous signal sequence selectedpreferably is one that is recognized and processed through one of thestandard pathways available within the host cell. The S. cerevisiaealpha factor pre-pro signal has proven effective in the secretion of avariety of recombinant proteins from P. pastoris. Other yeast signalsequences include the alpha mating factor signal sequence, the invertasesignal sequence, and signal sequences derived from other secreted yeastpolypeptides. Additionally, these signal peptide sequences may beengineered to provide for enhanced secretion in diploid yeast expressionsystems. Other secretion signals of interest also include mammaliansignal sequences, which may be heterologous to the protein beingsecreted, or may be a native sequence for the protein being secreted.Signal sequences include pre-peptide sequences, and in some instancesmay include propeptide sequences. Many such signal sequences are knownin the art, including the signal sequences found on immunoglobulinchains, e.g., K28 preprotoxin sequence, PHA-E, FACE, human MCP-1, humanserum albumin signal sequences, human Ig heavy chain, human Ig lightchain, and the like. For example, see Hashimoto et. al. Protein Eng11(2) 75 (1998); and Kobayashi et. al. Therapeutic Apheresis 2(4) 257(1998).

Transcription may be increased by inserting a transcriptional activatorsequence into the vector. These activators are cis-acting elements ofDNA, usually about from 10 to 300 bp, which act on a promoter toincrease its transcription. Transcriptional enhancers are relativelyorientation and position independent, having been found 5′ and 3′ to thetranscription unit, within an intron, as well as within the codingsequence itself. The enhancer may be spliced into the expression vectorat a position 5′ or 3′ to the coding sequence, but is preferably locatedat a site 5′ from the promoter.

Expression vectors used in eukaryotic host cells may also containsequences necessary for the termination of transcription and forstabilizing the mRNA. Such sequences are commonly available from 3′ tothe translation termination codon, in untranslated regions of eukaryoticor viral DNAs or cDNAs. These regions contain nucleotide segmentstranscribed as polyadenylated fragments in the untranslated portion ofthe mRNA.

Construction of suitable vectors containing one or more of theabove-listed components employs standard ligation techniques orPCR/recombination methods. Isolated plasmids or DNA fragments arecleaved, tailored, and re-ligated in the form desired to generate theplasmids required or via recombination methods. For analysis to confirmcorrect sequences in plasmids constructed, the ligation mixtures areused to transform host cells, and successful transformants selected byantibiotic resistance (e.g. ampicillin or Zeocin) where appropriate.Plasmids from the transformants are prepared, analyzed by restrictionendonuclease digestion and/or sequenced.

As an alternative to restriction and ligation of fragments,recombination methods based on att sites and recombination enzymes maybe used to insert DNA sequences into a vector. Such methods aredescribed, for example, by Landy (1989) Ann. Rev. Biochem. 58:913-949;and are known to those of skill in the art. Such methods utilizeintermolecular DNA recombination that is mediated by a mixture of lambdaand E. coli-encoded recombination proteins. Recombination occurs betweenspecific attachment (att) sites on the interacting DNA molecules. For adescription of att sites see Weisberg and Landy (1983) Site-SpecificRecombination in Phage Lambda, in Lambda II, Weisberg, ed. (Cold SpringHarbor, N.Y.: Cold Spring Harbor Press), pp. 211-250. The DNA segmentsflanking the recombination sites are switched, such that afterrecombination, the att sites are hybrid sequences comprised of sequencesdonated by each parental vector. The recombination can occur betweenDNAs of any topology.

Att sites may be introduced into a sequence of interest by ligating thesequence of interest into an appropriate vector; generating a PCRproduct containing att B sites through the use of specific primers;generating a cDNA library cloned into an appropriate vector containingatt sites; and the like.

Folding, as used herein, refers to the three-dimensional structure ofpolypeptides and proteins, where interactions between amino acidresidues act to stabilize the structure. While non-covalent interactionsare important in determining structure, usually the proteins of interestwill have intra- and/or intermolecular covalent disulfide bonds formedby two cysteine residues. For naturally occurring proteins andpolypeptides or derivatives and variants thereof, the proper folding istypically the arrangement that results in optimal biological activity,and can conveniently be monitored by assays for activity, e.g. ligandbinding, enzymatic activity, etc.

In some instances, for example where the desired product is of syntheticorigin, assays based on biological activity will be less meaningful. Theproper folding of such molecules may be determined on the basis ofphysical properties, energetic considerations, modeling studies, and thelike.

The expression host may be further modified by the introduction ofsequences encoding one or more enzymes that enhance folding anddisulfide bond formation, i.e. foldases, chaperonins, etc. Suchsequences may be constitutively or inducibly expressed in the yeast hostcell, using vectors, markers, etc. as known in the art. Preferably thesequences, including transcriptional regulatory elements sufficient forthe desired pattern of expression, are stably integrated in the yeastgenome through a targeted methodology.

For example, the eukaryotic PDI is not only an efficient catalyst ofprotein cysteine oxidation and disulfide bond isomerization, but alsoexhibits chaperone activity. Co-expression of PDI can facilitate theproduction of active proteins having multiple disulfide bonds. Also ofinterest is the expression of BIP (immunoglobulin heavy chain bindingprotein); cyclophilin; and the like. In one embodiment of the invention,each of the haploid parental strains expresses a distinct foldingenzyme, e.g. one strain may express BIP, and the other strain mayexpress PDI or combinations thereof.

The terms “desired protein” or “target protein” are used interchangeablyand refer generally to a humanized antibody or a binding portion thereofdescribed herein. The term “antibody” is intended to include anypolypeptide chain-containing molecular structure with a specific shapethat fits to and recognizes an epitope, where one or more non-covalentbinding interactions stabilize the complex between the molecularstructure and the epitope. The archetypal antibody molecule is theimmunoglobulin, and all types of immunoglobulins, IgG, IgM, IgA, IgE,IgD, etc., from all sources, e.g. human, rodent, rabbit, cow, sheep,pig, dog, other mammals, chicken, other avians, etc., are considered tobe “antibodies.” A preferred source for producing antibodies useful asstarting material according to the invention is rabbits. Numerousantibody coding sequences have been described; and others may be raisedby methods well-known in the art. Examples thereof include chimericantibodies, human antibodies and other non-human mammalian antibodies,humanized antibodies, single chain antibodies such as scFvs,camelbodies, nanobodies, IgNAR (single-chain antibodies derived fromsharks), small-modular immunopharmaceuticals (SMIPs), and antibodyfragments such as Fabs, Fab′, F(ab′)₂ and the like. See Streltsov V A,et al., Structure of a shark IgNAR antibody variable domain and modelingof an early-developmental isotype, Protein Sci. 2005 November;14(11):2901-9. Epub 2005 Sep. 30; Greenberg A S, et al., A new antigenreceptor gene family that undergoes rearrangement and extensive somaticdiversification in sharks, Nature. 1995 Mar. 9; 374(6518):168-73;Nuttall S D, et al., Isolation of the new antigen receptor fromwobbegong sharks, and use as a scaffold for the display of protein looplibraries, Mol Immunol. 2001 August; 38(4):313-26; Hamers-Casterman C,et al., Naturally occurring antibodies devoid of light chains, Nature.1993 Jun. 3; 363(6428):446-8; Gill D S, et al., Biopharmaceutical drugdiscovery using novel protein scaffolds, Curr Opin Biotechnol. 2006December; 17(6):653-8. Epub 2006 Oct. 19.

For example, antibodies or antigen binding fragments may be produced bygenetic engineering. In this technique, as with other methods,antibody-producing cells are sensitized to the desired antigen orimmunogen. The messenger RNA isolated from antibody producing cells isused as a template to make cDNA using PCR amplification. A library ofvectors, each containing one heavy chain gene and one light chain generetaining the initial antigen specificity, is produced by insertion ofappropriate sections of the amplified immunoglobulin cDNA into theexpression vectors. A combinatorial library is constructed by combiningthe heavy chain gene library with the light chain gene library. Thisresults in a library of clones which co-express a heavy and light chain(resembling the Fab fragment or antigen binding fragment of an antibodymolecule). The vectors that carry these genes are co-transfected into ahost cell. When antibody gene synthesis is induced in the transfectedhost, the heavy and light chain proteins self-assemble to produce activeantibodies that can be detected by screening with the antigen orimmunogen.

Antibody coding sequences of interest include those encoded by nativesequences, as well as nucleic acids that, by virtue of the degeneracy ofthe genetic code, are not identical in sequence to the disclosed nucleicacids, and variants thereof. Variant polypeptides can include amino acid(aa) substitutions, additions or deletions. The amino acid substitutionscan be conservative amino acid substitutions or substitutions toeliminate non-essential amino acids, such as to alter a glycosylationsite, or to minimize misfolding by substitution or deletion of one ormore cysteine residues that are not necessary for function. Variants canbe designed so as to retain or have enhanced biological activity of aparticular region of the protein (e.g., a functional domain, catalyticamino acid residues, etc). Variants also include fragments of thepolypeptides disclosed herein, particularly biologically activefragments and/or fragments corresponding to functional domains.Techniques for in vitro mutagenesis of cloned genes are known. Alsoincluded in the subject invention are polypeptides that have beenmodified using ordinary molecular biological techniques so as to improvetheir resistance to proteolytic degradation or to optimize solubilityproperties or to render them more suitable as a therapeutic agent.

Chimeric antibodies may be made by recombinant means by combining thevariable light and heavy chain regions (V_(L) and V_(H)), obtained fromantibody producing cells of one species with the constant light andheavy chain regions from another. Typically chimeric antibodies utilizerodent or rabbit variable regions and human constant regions, in orderto produce an antibody with predominantly human domains. The productionof such chimeric antibodies is well known in the art, and may beachieved by standard means (as described, e.g., in U.S. Pat. No.5,624,659, incorporated herein by reference in its entirety). It isfurther contemplated that the human constant regions of chimericantibodies of the invention may be selected from IgGI, IgG2, IgG3, IgG4,IgG5, IgG6, IgG7, IgG8, IgG9, IgGI0, IgGII, IgGI2, IgGI3, IgGI4, IgGI5,IgGI6, IgGI7, IgGI8 or IgGI9 constant regions.

Humanized antibodies are engineered to contain even more human-likeimmunoglobulin domains, and incorporate only thecomplementarity-determining regions of the animal-derived antibody. Thisis accomplished by carefully examining the sequence of thehyper-variable loops of the variable regions of the monoclonal antibody,and fitting them to the structure of the human antibody chains. Althoughfacially complex, the process is straightforward in practice. See, e.g.,U.S. Pat. No. 6,187,287, incorporated fully herein by reference.

In addition to entire immunoglobulins (or their recombinantcounterparts), immunoglobulin fragments comprising the epitope bindingsite (e.g., Fab′, F(ab′)₂, or other fragments) may be synthesized.“Fragment,” or minimal immunoglobulins may be designed utilizingrecombinant immunoglobulin techniques. For instance “Fv” immunoglobulinsfor use in the present invention may be produced by synthesizing a fusedvariable light chain region and a variable heavy chain region.Combinations of antibodies are also of interest, e.g. diabodies, whichcomprise two distinct Fv specificities. In another embodiment of theinvention, SMIPs (small molecule immunopharmaceuticals), camelbodies,nanobodies, and IgNAR are encompassed by immunoglobulin fragments.

Immunoglobulins and fragments thereof may be modifiedpost-translationally, e.g. to add effector moieties such as chemicallinkers, detectable moieties, such as fluorescent dyes, enzymes, toxins,substrates, bioluminescent materials, radioactive materials,chemiluminescent moieties and the like, or specific binding moieties,such as streptavidin, avidin, or biotin, and the like may be utilized inthe methods and compositions of the present invention. Examples ofadditional effector molecules are provided infra.

The term “polyploid yeast that stably expresses or expresses a desiredsecreted heterologous polypeptide for prolonged time” refers to a yeastculture that secretes said polypeptide for at least several days to aweek, more preferably at least a month, still more preferably at least1-6 months, and even more preferably for more than a year at thresholdexpression levels, typically at least 10-25 mg/liter and preferablysubstantially greater.

The term “polyploidal yeast culture that secretes desired amounts ofrecombinant polypeptide” refers to cultures that stably or for prolongedperiods secrete at least 10-25 mg/liter of heterologous polypeptide,more preferably at least 50-500 mg/liter, and most preferably 500-1000mg/liter or more.

A polynucleotide sequence “corresponds” to a polypeptide sequence iftranslation of the polynucleotide sequence in accordance with thegenetic code yields the polypeptide sequence (i.e., the polynucleotidesequence “encodes” the polypeptide sequence), one polynucleotidesequence “corresponds” to another polynucleotide sequence if the twosequences encode the same polypeptide sequence.

A “heterologous” region or domain of a DNA construct is an identifiablesegment of DNA within a larger DNA molecule that is not found inassociation with the larger molecule in nature. Thus, when theheterologous region encodes a mammalian gene, the gene will usually beflanked by DNA that does not flank the mammalian genomic DNA in thegenome of the source organism. Another example of a heterologous regionis a construct where the coding sequence itself is not found in nature(e.g., a cDNA where the genomic coding sequence contains introns, orsynthetic sequences having codons different than the native gene).Allelic variations or naturally-occurring mutational events do not giverise to a heterologous region of DNA as defined herein.

A “coding sequence” is an in-frame sequence of codons that (in view ofthe genetic code) correspond to or encode a protein or peptide sequence.Two coding sequences correspond to each other if the sequences or theircomplementary sequences encode the same amino acid sequences. A codingsequence in association with appropriate regulatory sequences may betranscribed and translated into a polypeptide. A polyadenylation signaland transcription termination sequence will usually be located 3′ to thecoding sequence. A “promoter sequence” is a DNA regulatory regioncapable of binding RNA polymerase in a cell and initiating transcriptionof a downstream (3′ direction) coding sequence. Promoter sequencestypically contain additional sites for binding of regulatory molecules(e.g., transcription factors) which affect the transcription of thecoding sequence. A coding sequence is “under the control” of thepromoter sequence or “operatively linked” to the promoter when RNApolymerase binds the promoter sequence in a cell and transcribes thecoding sequence into mRNA, which is then in turn translated into theprotein encoded by the coding sequence.

Vectors are used to introduce a foreign substance, such as DNA, RNA orprotein, into an organism or host cell. Typical vectors includerecombinant viruses (for polynucleotides) and liposomes (forpolypeptides). A “DNA vector” is a replicon, such as plasmid, phage orcosmid, to which another polynucleotide segment may be attached so as tobring about the replication of the attached segment. An “expressionvector” is a DNA vector which contains regulatory sequences which willdirect polypeptide synthesis by an appropriate host cell. This usuallymeans a promoter to bind RNA polymerase and initiate transcription ofmRNA, as well as ribosome binding sites and initiation signals to directtranslation of the mRNA into a polypeptide(s). Incorporation of apolynucleotide sequence into an expression vector at the proper site andin correct reading frame, followed by transformation of an appropriatehost cell by the vector, enables the production of a polypepide encodedby said polynucleotide sequence.

“Amplification” of polynucleotide sequences is the in vitro productionof multiple copies of a particular nucleic acid sequence. The amplifiedsequence is usually in the form of DNA. A variety of techniques forcarrying out such amplification are described in a review article by VanBrunt (1990, Bio/Technol., 8(4):291-294). Polymerase chain reaction orPCR is a prototype of nucleic acid amplification, and use of PCR hereinshould be considered exemplary of other suitable amplificationtechniques.

The general structure of antibodies in vertebrates now is wellunderstood (Edelman, G. M., Ann. N.Y. Acad. Sci., 190: 5 (1971)).Antibodies consist of two identical light polypeptide chains ofmolecular weight approximately 23,000 daltons (the “light chain”), andtwo identical heavy chains of molecular weight 53,000-70,000 (the “heavychain”). The four chains are joined by disulfide bonds in a “Y”configuration wherein the light chains bracket the heavy chains startingat the mouth of the “Y” configuration. The “branch” portion of the “Y”configuration is designated the F_(ab) region; the stem portion of the“Y” configuration is designated the Fc region. The amino acid sequenceorientation runs from the N-terminal end at the top of the “Y”configuration to the C-terminal end at the bottom of each chain. TheN-terminal end possesses the variable region having specificity for theantigen that elicited it, and is approximately 100 amino acids inlength, there being slight variations between light and heavy chain andfrom antibody to antibody.

The variable region is linked in each chain to a constant region thatextends the remaining length of the chain and that within a particularclass of antibody does not vary with the specificity of the antibody(i.e., the antigen eliciting it). There are five known major classes ofconstant regions that determine the class of the immunoglobulin molecule(IgG, IgM, IgA, IgD, and IgE corresponding to γ, μ, α, δ, and ε (gamma,mu, alpha, delta, or epsilon) heavy chain constant regions). Theconstant region or class determines subsequent effector function of theantibody, including activation of complement (Kabat, E. A., StructuralConcepts in Immunology and Immunochemistry, 2nd Ed., p. 413-436, Holt,Rinehart, Winston (1976)), and other cellular responses (Andrews, D. W.,et al., Clinical Immunobiology, pp 1-18, W. B. Sanders (1980); Kohl, S.,et al., Immunology, 48: 187 (1983)); while the variable regiondetermines the antigen with which it will react. Light chains areclassified as either κ (kappa) or λ (lambda). Each heavy chain class canbe paired with either kappa or lambda light chain. The light and heavychains are covalently bonded to each other, and the “tail” portions ofthe two heavy chains are bonded to each other by covalent disulfidelinkages when the immunoglobulins are generated either by hybridomas orby B cells.

The expression “variable region” or “VR” refers to the domains withineach pair of light and heavy chains in an antibody that are involveddirectly in binding the antibody to the antigen. Each heavy chain has atone end a variable domain (V_(H)) followed by a number of constantdomains. Each light chain has a variable domain (V_(L)) at one end and aconstant domain at its other end; the constant domain of the light chainis aligned with the first constant domain of the heavy chain, and thelight chain variable domain is aligned with the variable domain of theheavy chain.

The expressions “complementarity determining region,” “hypervariableregion,” or “CDR” refer to one or more of the hyper-variable orcomplementarity determining regions (CDRs) found in the variable regionsof light or heavy chains of an antibody (See Kabat, E. A. et al.,Sequences of Proteins of Immunological Interest, National Institutes ofHealth, Bethesda, Md., (1987)). These expressions include thehypervariable regions as defined by Kabat et al. (“Sequences of Proteinsof Immunological Interest,” Kabat E., et al., US Dept. of Health andHuman Services, 1983) or the hypervariable loops in 3-dimensionalstructures of antibodies (Chothia and Lesk, J Mol. Biol. 196 901-917(1987)). The CDRs in each chain are held in close proximity by frameworkregions and, with the CDRs from the other chain, contribute to theformation of the antigen binding site. Within the CDRs there are selectamino acids that have been described as the selectivity determiningregions (SDRs) which represent the critical contact residues used by theCDR in the antibody-antigen interaction (Kashmiri, S., Methods, 36:25-34(2005)).

The expressions “framework region” or “FR” refer to one or more of theframework regions within the variable regions of the light and heavychains of an antibody (See Kabat, E. A. et al., Sequences of Proteins ofImmunological Interest, National Institutes of Health, Bethesda, Md.,(1987)). These expressions include those amino acid sequence regionsinterposed between the CDRs within the variable regions of the light andheavy chains of an antibody.

Anti-IL-6 Antibodies and Binding Fragments Thereof

The invention includes antibodies having binding specificity to IL-6 andpossessing a variable light chain sequence comprising the sequence setforth below:

(SEQ ID NO: 2) MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKPGQRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSLRNIDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQL KSGTASVVCLLNN.

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 3) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYVTWVRQAPGKGLEWIGIIYGSDETAYATWAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNLWGQGTLVTVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 4; SEQ ID NO: 5; and SEQ ID NO:6 which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 2, and/or one or more of the polypeptide sequences of SEQ ID NO: 7;SEQ ID NO: 8; and SEQ ID NO: 9 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 3, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 4; SEQ ID NO: 5; and SEQ ID NO: 6which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 2, and/or one or more of the polypeptide sequences of SEQ ID NO: 7;SEQ ID NO: 8; and SEQ ID NO: 9 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 3, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 2. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 3.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 4; SEQ ID NO:5; and SEQ ID NO: 6 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable light chainsequence of SEQ ID NO: 2.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 7; SEQ ID NO:8; and SEQ ID NO: 9 which correspond to the complementarity-determiningregions (CDRs, or hypervariable regions) of the variable heavy chainsequence of SEQ ID NO: 3.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 2; the variable heavy chain region of SEQ IDNO: 3; the complementarity-determining regions (SEQ ID NO: 4; SEQ ID NO:5; and SEQ ID NO: 6) of the variable light chain region of SEQ ID NO: 2;and the complementarity-determining regions (SEQ ID NO: 7; SEQ ID NO: 8;and SEQ ID NO: 9) of the variable heavy chain region of SEQ ID NO: 3.

The invention also contemplates variants wherein either of the heavychain polypeptide sequences of SEQ ID NO: 18 or SEQ ID NO: 19 issubstituted for the heavy chain polypeptide sequence of SEQ ID NO: 3;the light chain polypeptide sequence of SEQ ID NO: 20 is substituted forthe light chain polypeptide sequence of SEQ ID NO: 2; and the heavychain CDR sequence of SEQ ID NO: 120 is substituted for the heavy chainCDR sequence of SEQ ID NO: 8.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb1, comprising SEQ ID NO: 2 and SEQ ID NO: 3, or the alternative SEQ IDNOs set forth in the preceding paragraph, and having at least one of thebiological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 21) MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTINCQASETIYSWLSWYQQKPGQPPKLLIYQASDLASGVPSRFSGSGAGTEYTLTISGVQCDDAATYYCQQGYSGSNVDNVFGGGTEVVVKRTVAAPSVFIFPPSDEQL KSGTASVVCLLNNFY

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 22) METGLRWLLLVAVLKGVQCQEQLKESGGRLVTPGTPLTLTCTASGFSLNDHAMGWVRQAPGKGLEYIGFINSGGSARYASWAEGRFTISRTSTTVDLKMTSLTTEDTATYFCVRGGAVWSIHSFDPWGPGTLVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 23; SEQ ID NO: 24; and SEQ IDNO: 25 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 21, and/or one or more of the polypeptide sequences of SEQ IDNO: 26; SEQ ID NO: 27; and SEQ ID NO: 28 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 22, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 23; SEQ ID NO: 24; and SEQ ID NO: 25which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 21, and/or one or more of the polypeptide sequences of SEQ ID NO:26; SEQ ID NO: 27; and SEQ ID NO: 28 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 22, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 21. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 22.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 23; SEQ IDNO: 24; and SEQ ID NO: 25 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 21.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 26; SEQ IDNO: 27; and SEQ ID NO: 28 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 22.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 21; the variable heavy chain region of SEQ IDNO: 22; the complementarity-determining regions (SEQ ID NO: 23; SEQ IDNO: 24; and SEQ ID NO: 25) of the variable light chain region of SEQ IDNO: 21; and the complementarity-determining regions (SEQ ID NO: 26; SEQID NO: 27; and SEQ ID NO: 28) of the variable heavy chain region of SEQID NO: 22.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb2, comprising SEQ ID NO: 21 and SEQ ID NO: 22, and having at least oneof the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 37) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQASQSVYDNNYLSWFQQKPGQPPKLLIYGASTLASGVPSRFVGSGSGTQFTLTITDVQCDDAATYYCAGVYDDDSDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNN

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 38) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSVYYMNWVRQAPGKGLEWIGFITMSDNINYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARSRGWGTMGRLDLWGPGTLVTVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 39; SEQ ID NO: 40; and SEQ IDNO: 41 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 37, and/or one or more of the polypeptide sequences of SEQ IDNO: 42; SEQ ID NO: 43; and SEQ ID NO: 44 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 38, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 39; SEQ ID NO: 40; and SEQ ID NO: 41which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 37, and/or one or more of the polypeptide sequences of SEQ ID NO:42; SEQ ID NO: 43; and SEQ ID NO: 44 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 38, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 37. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 38.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 39; SEQ IDNO: 40; and SEQ ID NO: 41 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 37.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 42; SEQ IDNO: 43; and SEQ ID NO: 44 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 38.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 37; the variable heavy chain region of SEQ IDNO: 38; the complementarity-determining regions (SEQ ID NO: 39; SEQ IDNO: 40; and SEQ ID NO: 41) of the variable light chain region of SEQ IDNO: 37; and the complementarity-determining regions (SEQ ID NO: 42; SEQID NO: 43; and SEQ ID NO: 44) of the variable heavy chain region of SEQID NO: 38.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb3, comprising SEQ ID NO: 37 and SEQ ID NO: 38, and having at least oneof the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 53) MDTRAPTQLLGLLLLWLPGAICDPVLTQTPSPVSAPVGGTVSISCQASQSVYENNYLSWFQQKPGQPPKLLIYGASTLDSGVPSRFKGSGSGTQFTLTITDVQCDDAATYYCAGVYDDDSDDAFGGGTEVVVKRTVAAPSVFIFPPSDEQ LKSGTASVVCLLNN

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 54) METGLRWLLLVAVLKGVQCQEQLKESGGGLVTPGGTLTLTCTASGFSLNAYYMNWVRQAPGKGLEWIGFITLNNNVAYANWAKGRFTFSKTSTTVDLKMTSPTPEDTATYFCARSRGWGAMGRLDLWGHGTLVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; and SEQ IDNO: 57 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 53, and/or one or more of the polypeptide sequences of SEQ IDNO: 58; SEQ ID NO: 59; and SEQ ID NO: 60 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 54, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 55; SEQ ID NO: 56; and SEQ ID NO: 57which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 53, and/or one or more of the polypeptide sequences of SEQ ID NO:58; SEQ ID NO: 59; and SEQ ID NO: 60 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 54, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 53. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 54.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 55; SEQ IDNO: 56; and SEQ ID NO: 57 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 53.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 58; SEQ IDNO: 59; and SEQ ID NO: 60 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 54.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 53; the variable heavy chain region of SEQ IDNO: 54; the complementarity-determining regions (SEQ ID NO: 55; SEQ IDNO: 56; and SEQ ID NO: 57) of the variable light chain region of SEQ IDNO: 53; and the complementarity-determining regions (SEQ ID NO: 58; SEQID NO: 59; and SEQ ID NO: 60) of the variable heavy chain region of SEQID NO: 54.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb4, comprising SEQ ID NO: 53 and SEQ ID NO: 54, and having at least oneof the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 69) MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSPVSAAVGGTVTINCQASQSVDDNNWLGWYQQKRGQPPKYLIYSASTLASGVPSRFKGSGSGTQFTLTISDLECDDAATYYCAGGFSGNIFAFGGGTEVVVKRTVAAPSVFIFPPSDEQL KSGTASVVCLLNNF

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 70) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYAMSWVRQAPGKGLEWIGIIGGFGTTYYATWAKGRFTISKTSTTVDLRITSPTTEDTATYFCARGGPGNGGDIWGQGTLVTVSSASTKGPSVFPLAPSSKS TSGGTAALGCLVKD.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 71; SEQ ID NO: 72; and SEQ IDNO: 73 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 69, and/or one or more of the polypeptide sequences of SEQ IDNO: 74; SEQ ID NO: 75; and SEQ ID NO: 76 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 70, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 71; SEQ ID NO: 72; and SEQ ID NO: 73which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 69, and/or one or more of the polypeptide sequences of SEQ ID NO:74; SEQ ID NO: 75; and SEQ ID NO: 76 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 70, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 69. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 70.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 71; SEQ IDNO: 72; and SEQ ID NO: 73 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 69.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 74; SEQ IDNO: 75; and SEQ ID NO: 76 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 70.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 69; the variable heavy chain region of SEQ IDNO: 70; the complementarity-determining regions (SEQ ID NO: 71; SEQ IDNO: 72; and SEQ ID NO: 73) of the variable light chain region of SEQ IDNO: 69; and the complementarity-determining regions (SEQ ID NO: 74; SEQID NO: 75; and SEQ ID NO: 76) of the variable heavy chain region of SEQID NO: 70.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb5, comprising SEQ ID NO: 69 and SEQ ID NO: 70, and having at least oneof the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 85) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSVPVGGTVTIKCQSSQSVYNNFLSWYQQKPGQPPKLLIYQASKLASGVPDRFSGSGSGTQFTLTISGVQCDDAATYYCLGGYDDDADNAFGGGTEVVVKRTVAAPSVFIFPPSDEQL KSGTASVVCLLNNF

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 86) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGIDLSDYAMSWVRQAPGKGLEWIGIIYAGSGSTWYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARDGYDDYGDFDRLDLWGPGTLVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKD.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 87; SEQ ID NO: 88; and SEQ IDNO: 89 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 85, and/or one or more of the polypeptide sequences of SEQ IDNO: 90; SEQ ID NO: 91; and SEQ ID NO: 92 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 86, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 87; SEQ ID NO: 88; and SEQ ID NO: 89which correspond to the complementarity-determining regions (CDRs, orhypervariable regions) of the variable light chain sequence of SEQ IDNO: 85, and/or one or more of the polypeptide sequences of SEQ ID NO:90; SEQ ID NO: 91; and SEQ ID NO: 92 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 86, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 85. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 86.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 87; SEQ IDNO: 88; and SEQ ID NO: 89 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 85.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 90; SEQ IDNO: 91; and SEQ ID NO: 92 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 86.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 85; the variable heavy chain region of SEQ IDNO: 86; the complementarity-determining regions (SEQ ID NO: 87; SEQ IDNO: 88; and SEQ ID NO: 89) of the variable light chain region of SEQ IDNO: 85; and the complementarity-determining regions (SEQ ID NO: 90; SEQID NO: 91; and SEQ ID NO: 92) of the variable heavy chain region of SEQID NO: 86.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb6, comprising SEQ ID NO: 85 and SEQ ID NO: 86, and having at least oneof the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 101) MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVSAAVGGTVTIKCQASQSINNELSWYQQKSGQRPKLLIYRASTLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSLRNIDNAFGGGTEVVVKRTVAAPSVFIFPPSDEQL KSGTASVVCLLNNF

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 102) METGLRWLLLVAVLSGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLSNYYMTWVRQAPGKGLEWIGMIYGSDETAYANWAIGRFTISKTSTTVDLKMTSLTAADTATYFCARDDSSDWDAKFNLWGQGTLVTVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVK.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 103; SEQ ID NO: 104; and SEQ IDNO: 105 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 101, and/or one or more of the polypeptide sequences of SEQID NO: 106; SEQ ID NO: 107; and SEQ ID NO: 108 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 102, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 103; SEQ ID NO: 104; and SEQ ID NO:105 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 101, and/or one or more of the polypeptide sequences of SEQ ID NO:106; SEQ ID NO: 107; and SEQ ID NO: 108 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 102, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 101. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 102.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 103; SEQ IDNO: 104; and SEQ ID NO: 105 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 101.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 106; SEQ IDNO: 107; and SEQ ID NO: 108 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 102.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 101; the variable heavy chain region of SEQID NO: 102; the complementarity-determining regions (SEQ ID NO: 103; SEQID NO: 104; and SEQ ID NO: 105) of the variable light chain region ofSEQ ID NO: 101; and the complementarity-determining regions (SEQ ID NO:106; SEQ ID NO: 107; and SEQ ID NO: 108) of the variable heavy chainregion of SEQ ID NO: 102.

The invention also contemplates variants wherein either of the heavychain polypeptide sequences of SEQ ID NO: 117 or SEQ ID NO: 118 issubstituted for the heavy chain polypeptide sequence of SEQ ID NO: 102;the light chain polypeptide sequence of SEQ ID NO: 119 is substitutedfor the light chain polypeptide sequence of SEQ ID NO: 101; and theheavy chain CDR sequence of SEQ ID NO: 121 is substituted for the heavychain CDR sequence of SEQ ID NO: 107.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb7, comprising SEQ ID NO: 101 and SEQ ID NO: 102, or the alternativeSEQ ID NOs set forth in the preceding paragraph, and having at least oneof the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 122) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVTISCQSSQSVGNNQDLSWFQQRPGQPPKLLIYEISKLESGVPSRFSGSGSGTHFTLTISGVQCDDAATYYCLGGYDDDADNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 123) METGLRWLLLVAVLKGVQCHSVEESGGRLVTPGTPLTLTCTVSGFSLSSRTMSWVRQAPGKGLEWIGYIWSGGSTYYATWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARLGDTGGHAYATRLNL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 124; SEQ ID NO: 125; and SEQ IDNO: 126 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 122, and/or one or more of the polypeptide sequences of SEQID NO: 127; SEQ ID NO: 128; and SEQ ID NO: 129 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 123, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 124; SEQ ID NO: 125; and SEQ ID NO:126 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 122, and/or one or more of the polypeptide sequences of SEQ ID NO:127; SEQ ID NO: 128; and SEQ ID NO: 129 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 123, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 122. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 123.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 124; SEQ IDNO: 125; and SEQ ID NO: 126 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 122.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 127; SEQ IDNO: 128; and SEQ ID NO: 129 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 123.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 122; the variable heavy chain region of SEQID NO: 123; the complementarity-determining regions (SEQ ID NO: 124; SEQID NO: 125; and SEQ ID NO: 126) of the variable light chain region ofSEQ ID NO: 122; and the complementarity-determining regions (SEQ ID NO:127; SEQ ID NO: 128; and SEQ ID NO: 129) of the variable heavy chainregion of SEQ ID NO: 123.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb8, comprising SEQ ID NO: 122 and SEQ ID NO: 123, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 138) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSSVSAAVGGTVSISCQSSQSVYSNKYLAWYQQKPGQPPKLLIYWTSKLASGAPSRFSGSGSGTQFTLTISGVQCDDAATYYCLGAYDDDADNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 139) METGLRWLLLVAVLKGVQCQSVEESGGRLVKPDETLTLTCTASGFSLEGGYMTWVRQAPGKGLEWIGISYDSGSTYYASWAKGRFTISKTSSTTVDLKMTSLTTEDTATYFCVRSLKYPTVTSDDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 140; SEQ ID NO: 141; and SEQ IDNO: 142 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 138, and/or one or more of the polypeptide sequences of SEQID NO: 143; SEQ ID NO: 144; and SEQ ID NO: 145 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 139, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 140; SEQ ID NO: 141; and SEQ ID NO:142 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 138, and/or one or more of the polypeptide sequences of SEQ ID NO:143; SEQ ID NO: 144; and SEQ ID NO: 145 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 139, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 138. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 139.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 140; SEQ IDNO: 141; and SEQ ID NO: 142 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 138.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 143; SEQ IDNO: 144; and SEQ ID NO: 145 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 139.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 138; the variable heavy chain region of SEQID NO: 139; the complementarity-determining regions (SEQ ID NO: 140; SEQID NO: 141; and SEQ ID NO: 142) of the variable light chain region ofSEQ ID NO: 138; and the complementarity-determining regions (SEQ ID NO:143; SEQ ID NO: 144; and SEQ ID NO: 145) of the variable heavy chainregion of SEQ ID NO: 139.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb9, comprising SEQ ID NO: 138 and SEQ ID NO: 139, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 154) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVTISCQSSQSVYNNNDLAWYQQKPGQPPKLLIYYASTLASGVPSRFKGSGSGTQFTLTISGVQCDDAAAYYCLGGYDDDADNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 155) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGLSLSSNTINWVRQAPGKGLEWIGYIWSGGSTYYASWVNGRFTISKTSTTVDLKITSPTTEDTATYFCARGGYASGGYPYATRLDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 156; SEQ ID NO: 157; and SEQ IDNO: 158 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 154, and/or one or more of the polypeptide sequences of SEQID NO: 159; SEQ ID NO: 160; and SEQ ID NO: 161 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 155, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 156; SEQ ID NO: 157; and SEQ ID NO:158 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 154, and/or one or more of the polypeptide sequences of SEQ ID NO:159; SEQ ID NO: 160; and SEQ ID NO: 161 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 155, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 154. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 155.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 156; SEQ IDNO: 157; and SEQ ID NO: 158 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 154.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 159; SEQ IDNO: 160; and SEQ ID NO: 161 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 155.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 154; the variable heavy chain region of SEQID NO: 155; the complementarity-determining regions (SEQ ID NO: 156; SEQID NO: 157; and SEQ ID NO: 158) of the variable light chain region ofSEQ ID NO: 154; and the complementarity-determining regions (SEQ ID NO:159; SEQ ID NO: 160; and SEQ ID NO: 161) of the variable heavy chainregion of SEQ ID NO: 155.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb10, comprising SEQ ID NO: 154 and SEQ ID NO: 155, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 170) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSSVSAAVGGTVTINCQSSQSVYNNDYLSWYQQRPGQRPKLLIYGASKLASGVPSRFKGSGSGKQFTLTISGVQCDDAATYYCLGDYDDDADNT

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 171) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFTLSTNYYLSWVRQAPGKGLEWIGIIYPSGNTYCAKWAKGRFTISKTSSTTVDLKMTSPTTEDTATYFCARNYGGDESL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 172; SEQ ID NO: 173; and SEQ IDNO: 174 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 170, and/or one or more of the polypeptide sequences of SEQID NO: 175; SEQ ID NO: 176; and SEQ ID NO: 177 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 171, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 172; SEQ ID NO: 173; and SEQ ID NO:174 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 170, and/or one or more of the polypeptide sequences of SEQ ID NO:175; SEQ ID NO: 176; and SEQ ID NO: 177 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 171, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 170. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 171.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 172; SEQ IDNO: 173; and SEQ ID NO: 174 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 170.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 175; SEQ IDNO: 176; and SEQ ID NO: 177 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 171.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 170; the variable heavy chain region of SEQID NO: 171; the complementarity-determining regions (SEQ ID NO: 172; SEQID NO: 173; and SEQ ID NO: 174) of the variable light chain region ofSEQ ID NO: 170; and the complementarity-determining regions (SEQ ID NO:175; SEQ ID NO: 176; and SEQ ID NO: 177) of the variable heavy chainregion of SEQ ID NO: 171.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb11, comprising SEQ ID NO: 170 and SEQ ID NO: 171, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 186) MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASETIGNALAWYQQKSGQPPKLLIYKASKLASGVPSRFKGSGSGTEYTLTISDL ECADAATYYCQWCYFGDSV

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 187) METGLRWLLLVTVLKGVQCQEQLVESGGGLVQPEGSLTLTCTASGFDFSSGYYMCWVRQAPGKGLEWIACIFTITTNTYYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYLCARGIYSDNNYYAL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 188; SEQ ID NO: 189; and SEQ IDNO: 190 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 186, and/or one or more of the polypeptide sequences of SEQID NO: 191; SEQ ID NO: 192; and SEQ ID NO: 193 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 187, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 188; SEQ ID NO: 189; and SEQ ID NO:190 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 186, and/or one or more of the polypeptide sequences of SEQ ID NO:191; SEQ ID NO: 192; and SEQ ID NO: 193 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 187, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 186. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 187.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 188; SEQ IDNO: 189; and SEQ ID NO: 190 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 186.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 191; SEQ IDNO: 192; and SEQ ID NO: 193 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 187.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 186; the variable heavy chain region of SEQID NO: 187; the complementarity-determining regions (SEQ ID NO: 188; SEQID NO: 189; and SEQ ID NO: 190) of the variable light chain region ofSEQ ID NO: 186; and the complementarity-determining regions (SEQ ID NO:191; SEQ ID NO: 192; and SEQ ID NO: 193) of the variable heavy chainregion of SEQ ID NO: 187.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb12, comprising SEQ ID NO: 186 and SEQ ID NO: 187, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 202) MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASESIGNALAWYQQKPGQPPKLLIYKASTLASGVPSRFSGSGSGTEFTLTISGV QCADAAAYYCQWCYFGDSV

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 203) METGLRWLLLVAVLKGVQCQQQLVESGGGLVKPGASLTLTCKASGFSFSSGYYMCWVRQAPGKGLESIACIFTITDNTYYANWAKGRFTISKPSSPTVTLQMTSLTAADTATYFCARGIYSTDNYYAL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 204; SEQ ID NO: 205; and SEQ IDNO: 206 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 202, and/or one or more of the polypeptide sequences of SEQID NO: 207; SEQ ID NO: 208; and SEQ ID NO: 209 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 203, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 204; SEQ ID NO: 205; and SEQ ID NO:206 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 202, and/or one or more of the polypeptide sequences of SEQ ID NO:207; SEQ ID NO: 208; and SEQ ID NO: 209 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 203, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 202. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 203.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 204; SEQ IDNO: 205; and SEQ ID NO: 206 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 202.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 207; SEQ IDNO: 208; and SEQ ID NO: 209 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 203.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 202; the variable heavy chain region of SEQID NO: 203; the complementarity-determining regions (SEQ ID NO: 204; SEQID NO: 205; and SEQ ID NO: 206) of the variable light chain region ofSEQ ID NO: 202; and the complementarity-determining regions (SEQ ID NO:207; SEQ ID NO: 208; and SEQ ID NO: 209) of the variable heavy chainregion of SEQ ID NO: 203.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb13, comprising SEQ ID NO: 202 and SEQ ID NO: 203, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 218) MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASQSVSSYLNWYQQKPGQPPKLLIYRASTLESGVPSRFKGSGSGTEFTLTISDLECADAATYYCQCTYGTSSSYGAA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 219) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGISLSSNAISWVRQAPGKGLEWIGIISYSGTTYYASWAKGRFTISKTSSTTVDLKITSPTTEDTATYFCARDDPTTVMVMLIPFGAGMDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 220; SEQ ID NO: 221; and SEQ IDNO: 222 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 218, and/or one or more of the polypeptide sequences of SEQID NO: 223; SEQ ID NO: 224; and SEQ ID NO: 225 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 219, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 220; SEQ ID NO: 221; and SEQ ID NO:222 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 218, and/or one or more of the polypeptide sequences of SEQ ID NO:223; SEQ ID NO: 224; and SEQ ID NO: 225 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 219, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 218. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 219.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 220; SEQ IDNO: 221; and SEQ ID NO: 222 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 218.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 223; SEQ IDNO: 224; and SEQ ID NO: 225 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 219.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 218; the variable heavy chain region of SEQID NO: 219; the complementarity-determining regions (SEQ ID NO: 220; SEQID NO: 221; and SEQ ID NO: 222) of the variable light chain region ofSEQ ID NO: 218; and the complementarity-determining regions (SEQ ID NO:223; SEQ ID NO: 224; and SEQ ID NO: 225) of the variable heavy chainregion of SEQ ID NO: 219.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb14, comprising SEQ ID NO: 218 and SEQ ID NO: 219, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 234) MDTRAPTQLLGLLLLWLPGATFAQVLTQTASPVSAAVGGTVTINCQASQSVYKNNYLSWYQQKPGQPPKGLIYSASTLDSGVPLRFSGSGSGTQFTLTISDVQCDDAATYYCLGSYDCSSGDCYA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 235) METGLRWLLLVAVLKGVQCQSLEESGGDLVKPEGSLTLTCTASGFSFSSYWMCWVRQAPGKGLEWIACIVTGNGNTYYANWAKGRFTISKTSSTTVTLQM TSLTAADTATYFCAKAYDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 236; SEQ ID NO: 237; and SEQ IDNO: 238 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 234, and/or one or more of the polypeptide sequences of SEQID NO: 239; SEQ ID NO: 240; and SEQ ID NO: 241 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 235, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 236; SEQ ID NO: 237; and SEQ ID NO:238 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 234, and/or one or more of the polypeptide sequences of SEQ ID NO:239; SEQ ID NO: 240; and SEQ ID NO: 241 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 235, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 234. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 235.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 236; SEQ IDNO: 237; and SEQ ID NO: 238 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 234.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 239; SEQ IDNO: 240; and SEQ ID NO: 241 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 235.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 234; the variable heavy chain region of SEQID NO: 235; the complementarity-determining regions (SEQ ID NO: 236; SEQID NO: 237; and SEQ ID NO: 238) of the variable light chain region ofSEQ ID NO: 234; and the complementarity-determining regions (SEQ ID NO:239; SEQ ID NO: 240; and SEQ ID NO: 241) of the variable heavy chainregion of SEQ ID NO: 235.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb15, comprising SEQ ID NO: 234 and SEQ ID NO: 235, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 250) MDTRAPTQLLGLLLLWLPGSTFAAVLTQTPSPVSAAVGGTVSISCQASQSVYDNNYLSWYQQKPGQPPKLLIYGASTLASGVPSRFKGTGSGTQFTLTITDVQCDDAATYYCAGVFNDDSDDA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 251) METGLRWLLLVAVPKGVQCQSLEESGGRLVTPGTPLTLTCTLSGFSLSAYYMSWVRQAPGKGLEWIGFITLSDHISYARWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARSRGWGAMGRLDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 252; SEQ ID NO: 253; and SEQ IDNO: 254 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 250, and/or one or more of the polypeptide sequences of SEQID NO: 255; SEQ ID NO: 256; and SEQ ID NO: 257 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 251, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 252; SEQ ID NO: 253; and SEQ ID NO:254 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 250, and/or one or more of the polypeptide sequences of SEQ ID NO:255; SEQ ID NO: 256; and SEQ ID NO: 257 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 251, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 250. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 251.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 252; SEQ IDNO: 253; and SEQ ID NO: 254 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 250.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 255; SEQ IDNO: 256; and SEQ ID NO: 257 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 251.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 250; the variable heavy chain region of SEQID NO: 251; the complementarity-determining regions (SEQ ID NO: 252; SEQID NO: 253; and SEQ ID NO: 254) of the variable light chain region ofSEQ ID NO: 250; and the complementarity-determining regions (SEQ ID NO:255; SEQ ID NO: 256; and SEQ ID NO: 257) of the variable heavy chainregion of SEQ ID NO: 251.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb16, comprising SEQ ID NO: 250 and SEQ ID NO: 251, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 266) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVTISCQASQSVYNNKNLAWYQQKSGQPPKLLIYWASTLASGVSSRFSGSGSGTQFTLTVSGVQCDDAATYYCLGVFDDDADNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 267) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTASGFSLSSYSMTWVRQAPGKGLEYIGVIGTSGSTYYATWAKGRFTISRTSTTVALKITSPTTEDTATYFCVRSLSSITFL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 268; SEQ ID NO: 269; and SEQ IDNO: 270 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 266, and/or one or more of the polypeptide sequences of SEQID NO: 271; SEQ ID NO: 272; and SEQ ID NO: 273 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 267, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 268; SEQ ID NO: 269; and SEQ ID NO:270 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 266, and/or one or more of the polypeptide sequences of SEQ ID NO:271; SEQ ID NO: 272; and SEQ ID NO: 273 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 267, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 266. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 267.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 268; SEQ IDNO: 269; and SEQ ID NO: 270 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 266.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 271; SEQ IDNO: 272; and SEQ ID NO: 273 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 267.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 266; the variable heavy chain region of SEQID NO: 267; the complementarity-determining regions (SEQ ID NO: 268; SEQID NO: 269; and SEQ ID NO: 270) of the variable light chain region ofSEQ ID NO: 266; and the complementarity-determining regions (SEQ ID NO:271; SEQ ID NO: 272; and SEQ ID NO: 273) of the variable heavy chainregion of SEQ ID NO: 267.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb17, comprising SEQ ID NO: 266 and SEQ ID NO: 267, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 282) MDTRAPTQLLGLLLLWLPGARCAFELTQTPASVEAAVGGTVTINCQASQNIYRYLAWYQQKPGQPPKFLIYLASTLASGVPSRFKGSGSGTEFTLTISDL ECADAATYYCQSYYSSNSVA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 283) METGLRWLLLVAVLKGVQCQEQLVESGGDLVQPEGSLTLTCTASELDFSSGYWICWVRQVPGKGLEWIGCIYTGSSGSTFYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCARGYSGFGYFKL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 284; SEQ ID NO: 285; and SEQ IDNO: 286 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 282, and/or one or more of the polypeptide sequences of SEQID NO: 287; SEQ ID NO: 288; and SEQ ID NO: 289 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 283, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 284; SEQ ID NO: 285; and SEQ ID NO:286 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 282, and/or one or more of the polypeptide sequences of SEQ ID NO:287; SEQ ID NO: 288; and SEQ ID NO: 289 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 283, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 282. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 283.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 284; SEQ IDNO: 285; and SEQ ID NO: 286 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 282.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 287; SEQ IDNO: 288; and SEQ ID NO: 289 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 283.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 282; the variable heavy chain region of SEQID NO: 283; the complementarity-determining regions (SEQ ID NO: 284; SEQID NO: 285; and SEQ ID NO: 286) of the variable light chain region ofSEQ ID NO: 282; and the complementarity-determining regions (SEQ ID NO:287; SEQ ID NO: 288; and SEQ ID NO: 289) of the variable heavy chainregion of SEQ ID NO: 283.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb18, comprising SEQ ID NO: 282 and SEQ ID NO: 283, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 298) MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQASEDIYRLLAWYQQKPGQPPKLLIYDSSDLASGVPSRFKGSGSGTEFTLAISGVQCDDAATYYCQQAWSYSDIDNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 299) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTASGFSLSSYYMSWVRQAPGKGLEWIGIITTSGNTFYASWAKGRLTISRTSTTVDLKITSPTTEDTATYFCARTSDIFYYRNL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 300; SEQ ID NO: 301; and SEQ IDNO: 302 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 298, and/or one or more of the polypeptide sequences of SEQID NO: 303; SEQ ID NO: 304; and SEQ ID NO: 305 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 299, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 300; SEQ ID NO: 301; and SEQ ID NO:302 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 298, and/or one or more of the polypeptide sequences of SEQ ID NO:303; SEQ ID NO: 304; and SEQ ID NO: 305 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 299, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 298. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 299.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 300; SEQ IDNO: 301; and SEQ ID NO: 302 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 298.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 303; SEQ IDNO: 304; and SEQ ID NO: 305 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 299.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 298; the variable heavy chain region of SEQID NO: 299; the complementarity-determining regions (SEQ ID NO: 300; SEQID NO: 301; and SEQ ID NO: 302) of the variable light chain region ofSEQ ID NO: 298; and the complementarity-determining regions (SEQ ID NO:303; SEQ ID NO: 304; and SEQ ID NO: 305) of the variable heavy chainregion of SEQ ID NO: 299.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb19, comprising SEQ ID NO: 298 and SEQ ID NO: 299, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 314) MDTRAPTQLLGLLLLWLPGATFAAVLTQTASPVSAAVGATVTINCQSSQSVYNDMDLAWFQQKPGQPPKLLIYSASTLASGVPSRFSGSGSGTEFTLTISGVQCDDAATYYCLGAFDDDADNT

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 315) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLTRHAITWVRQAPGKGLEWIGCIWSGGSTYYATWAKGRFTISKTSTTVDLRITSPTTEDTATYFCARVIGDTAGYAYFTGLDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 316; SEQ ID NO: 317; and SEQ IDNO: 318 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 314, and/or one or more of the polypeptide sequences of SEQID NO: 319; SEQ ID NO: 320; and SEQ ID NO: 321 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 315, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 316; SEQ ID NO: 317; and SEQ ID NO:318 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 314, and/or one or more of the polypeptide sequences of SEQ ID NO:319; SEQ ID NO: 320; and SEQ ID NO: 321 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 315, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 314. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 315.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 316; SEQ IDNO: 317; and SEQ ID NO: 318 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 314.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 319; SEQ IDNO: 320; and SEQ ID NO: 321 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 315.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 314; the variable heavy chain region of SEQID NO: 315; the complementarity-determining regions (SEQ ID NO: 316; SEQID NO: 317; and SEQ ID NO: 318) of the variable light chain region ofSEQ ID NO: 314; and the complementarity-determining regions (SEQ ID NO:319; SEQ ID NO: 320; and SEQ ID NO: 321) of the variable heavy chainregion of SEQ ID NO: 315.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb20, comprising SEQ ID NO: 314 and SEQ ID NO: 315, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 330) MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQASQSVYNWLSWYQQKPGQPPKLLIYTASSLASGVPSRFSGSGSGTEFTLTISGV ECADAATYYCQQGYTSDVDNV

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 331) METGLRWLLLVAVLKGVQCQSLEEAGGRLVTPGTPLTLTCTVSGIDLSSYAMGWVRQAPGKGLEYIGIISSSGSTYYATWAKGRFTISQASSTTVDLKITSPTTEDSATYFCARGGAGSGGVWLLDGFDP.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 332; SEQ ID NO: 333; and SEQ IDNO: 334 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 330, and/or one or more of the polypeptide sequences of SEQID NO: 335; SEQ ID NO: 336; and SEQ ID NO: 337 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 331, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 332; SEQ ID NO: 333; and SEQ ID NO:334 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 330, and/or one or more of the polypeptide sequences of SEQ ID NO:335; SEQ ID NO: 336; and SEQ ID NO: 337 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 331, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 330. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 331.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 332; SEQ IDNO: 333; and SEQ ID NO: 334 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 330.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 335; SEQ IDNO: 336; and SEQ ID NO: 337 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 331.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 330; the variable heavy chain region of SEQID NO: 331; the complementarity-determining regions (SEQ ID NO: 332; SEQID NO: 333; and SEQ ID NO: 334) of the variable light chain region ofSEQ ID NO: 330; and the complementarity-determining regions (SEQ ID NO:335; SEQ ID NO: 336; and SEQ ID NO: 337) of the variable heavy chainregion of SEQ ID NO: 331.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb21, comprising SEQ ID NO: 330 and SEQ ID NO: 331, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 346) MDTRAPTQLLGLLLLWLPGAKCADVVMTQTPASVSAAVGGTVTINCQASENIYNWLAWYQQKPGQPPKLLIYTVGDLASGVSSRFKGSGSGTEFTLTISDLECADAATYYCQQGYSSSYVDNV

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 347) METGLRWLLLVAVLKGVQCQEQLKESGGRLVTPGTPLTLTCTVSGFSLNDYAVGWFRQAPGKGLEWIGYIRSSGTTAYATWAKGRFTISATSTTVDLKITSPTTEDTATYFCARGGAGSSGVWILDGFAP.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 348; SEQ ID NO: 349; and SEQ IDNO: 350 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 346, and/or one or more of the polypeptide sequences of SEQID NO: 351; SEQ ID NO: 352; and SEQ ID NO: 353 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 347, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 348; SEQ ID NO: 349; and SEQ ID NO:350 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 346, and/or one or more of the polypeptide sequences of SEQ ID NO:351; SEQ ID NO: 352; and SEQ ID NO: 353 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 347, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 346. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 347.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 348; SEQ IDNO: 349; and SEQ ID NO: 350 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 346.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 351; SEQ IDNO: 352; and SEQ ID NO: 353 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 347.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 346; the variable heavy chain region of SEQID NO: 347; the complementarity-determining regions (SEQ ID NO: 348; SEQID NO: 349; and SEQ ID NO: 350) of the variable light chain region ofSEQ ID NO: 346; and the complementarity-determining regions (SEQ ID NO:351; SEQ ID NO: 352; and SEQ ID NO: 353) of the variable heavy chainregion of SEQ ID NO: 347.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb22, comprising SEQ ID NO: 346 and SEQ ID NO: 347, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 362) MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSSVSAAVGGTVTINCQASQSVYQNNYLSWFQQKPGQPPKLLIYGAATLASGVPSRFKGSGSGTQFTLTIS DLECDDAATYYCAGAYRDVDS

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 363) METGLRWLLLVAVLKGVQCQSLEESGGDLVKPGASLTLTCTASGFSFTSTYYIYWVRQAPGKGLEWIACIDAGSSGSTYYATWVNGRFTISKTSSTTVTLQMTSLTAADTATYFCAKWDYGGNVGWGYDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 364; SEQ ID NO: 365; and SEQ IDNO: 366 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 362, and/or one or more of the polypeptide sequences of SEQID NO: 367; SEQ ID NO: 368; and SEQ ID NO: 369 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 363, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 364; SEQ ID NO: 365; and SEQ ID NO:366 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 362, and/or one or more of the polypeptide sequences of SEQ ID NO:367; SEQ ID NO: 368; and SEQ ID NO: 369 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 363, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 362. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 363.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 364; SEQ IDNO: 365; and SEQ ID NO: 366 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 362.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 367; SEQ IDNO: 368; and SEQ ID NO: 369 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 363.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 362; the variable heavy chain region of SEQID NO: 363; the complementarity-determining regions (SEQ ID NO: 364; SEQID NO: 365; and SEQ ID NO: 366) of the variable light chain region ofSEQ ID NO: 362; and the complementarity-determining regions (SEQ ID NO:367; SEQ ID NO: 368; and SEQ ID NO: 369) of the variable heavy chainregion of SEQ ID NO: 363.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb23, comprising SEQ ID NO: 362 and SEQ ID NO: 363, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 378) MDTRAPTQLLGLLLLWLPGARCAFELTQTPSSVEAAVGGTVTIKCQASQSISSYLAWYQQKPGQPPKFLIYRASTLASGVPSRFKGSGSGTEFTLTISDL ECADAATYYCQSYYDSVSNP

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 379) METGLRWLLLVAVLKGVQCQSLEESGGDLVKPEGSLTLTCKASGLDLGTYWFMCWVRQAPGKGLEWIACIYTGSSGSTFYASWVNGRFTISKTSSTTVTLQMTSLTAADTATYFCARGYSGYGYFKL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 380; SEQ ID NO: 381; and SEQ IDNO: 382 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 378, and/or one or more of the polypeptide sequences of SEQID NO: 383; SEQ ID NO: 384; and SEQ ID NO: 385 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 379, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 380; SEQ ID NO: 381; and SEQ ID NO:382 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 378, and/or one or more of the polypeptide sequences of SEQ ID NO:383; SEQ ID NO: 384; and SEQ ID NO: 385 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 379, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 378. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 379.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 380; SEQ IDNO: 381; and SEQ ID NO: 382 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 378.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 383; SEQ IDNO: 384; and SEQ ID NO: 385 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 379.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 378; the variable heavy chain region of SEQID NO: 379; the complementarity-determining regions (SEQ ID NO: 380; SEQID NO: 381; and SEQ ID NO: 382) of the variable light chain region ofSEQ ID NO: 378; and the complementarity-determining regions (SEQ ID NO:383; SEQ ID NO: 384; and SEQ ID NO: 385) of the variable heavy chainregion of SEQ ID NO: 379.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb24, comprising SEQ ID NO: 378 and SEQ ID NO: 379, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 394) MDTRAPTQLLGLLLLWLPGVTFAIEMTQSPFSVSAAVGGTVSISCQASQSVYKNNQLSWYQQKSGQPPKLLIYGASALASGVPSRFKGSGSGTEFTLTISDVQCDDAATYYCAGAITGSIDTDG

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 395) METGLRWLLLVAVLKGVQCQSLEESGGDLVKPGASLTLTCTTSGFSFSSSYFICWVRQAPGKGLEWIACIYGGDGSTYYASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCAREWAYSQGYFGAFDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 396; SEQ ID NO: 397; and SEQ IDNO: 398 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 394, and/or one or more of the polypeptide sequences of SEQID NO: 399; SEQ ID NO: 400; and SEQ ID NO: 401 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 395, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 396; SEQ ID NO: 397; and SEQ ID NO:398 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 394, and/or one or more of the polypeptide sequences of SEQ ID NO:399; SEQ ID NO: 400; and SEQ ID NO: 401 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 395, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 394. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 395.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 396; SEQ IDNO: 397; and SEQ ID NO: 398 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 394.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 399; SEQ IDNO: 400; and SEQ ID NO: 401 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 395.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 394; the variable heavy chain region of SEQID NO: 395; the complementarity-determining regions (SEQ ID NO: 396; SEQID NO: 397; and SEQ ID NO: 398) of the variable light chain region ofSEQ ID NO: 394; and the complementarity-determining regions (SEQ ID NO:399; SEQ ID NO: 400; and SEQ ID NO: 401) of the variable heavy chainregion of SEQ ID NO: 395.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb25, comprising SEQ ID NO: 394 and SEQ ID NO: 395, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 410) MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVEAAVGGTVTIKCQASEDISSYLAWYQQKPGQPPKLLIYAASNLESGVSSRFKGSGSGTEYTLTISDLECADAATYYCQCTYGTISISDGNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 411) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMTWVRQAPGEGLEYIGFINPGGSAYYASWVKGRFTISKSSTTVDLKITSPTTEDTATYFCARVLIVSYGAFTI.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 412; SEQ ID NO: 413; and SEQ IDNO: 414 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 410, and/or one or more of the polypeptide sequences of SEQID NO: 415; SEQ ID NO: 416; and SEQ ID NO: 417 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 411, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 412; SEQ ID NO: 413; and SEQ ID NO:414 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 410, and/or one or more of the polypeptide sequences of SEQ ID NO:415; SEQ ID NO: 416; and SEQ ID NO: 417 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 411, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 410. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 411.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 412; SEQ IDNO: 413; and SEQ ID NO: 414 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 410.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 415; SEQ IDNO: 416; and SEQ ID NO: 417 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 411.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 410; the variable heavy chain region of SEQID NO: 411; the complementarity-determining regions (SEQ ID NO: 412; SEQID NO: 413; and SEQ ID NO: 414) of the variable light chain region ofSEQ ID NO: 410; and the complementarity-determining regions (SEQ ID NO:415; SEQ ID NO: 416; and SEQ ID NO: 417) of the variable heavy chainregion of SEQ ID NO: 411.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb26, comprising SEQ ID NO: 410 and SEQ ID NO: 411, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 426) MDTRAPTQLLGLLLLWLPGARCDVVMTQTPASVSAAVGGTVTIKCQASEDIESYLAWYQQKPGQPPKLLIYGASNLESGVSSRFKGSGSGTEFTLTISDLECADAATYYCQCTYGIISISDGNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 427) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMTWVRQAPGEGLEYIGFMNTGDNAYYASWAKGRFTISKTSTTVDLKITSPTTEDTATYFCARVLVVAYGAFNI.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 428; SEQ ID NO: 429; and SEQ IDNO: 430 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 426, and/or one or more of the polypeptide sequences of SEQID NO: 431; SEQ ID NO: 432; and SEQ ID NO: 433 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 427, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 428; SEQ ID NO: 429; and SEQ ID NO:430 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 426, and/or one or more of the polypeptide sequences of SEQ ID NO:431; SEQ ID NO: 432; and SEQ ID NO: 433 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 427, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 426. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 427.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 428; SEQ IDNO: 429; and SEQ ID NO: 430 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 426.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 431; SEQ IDNO: 432; and SEQ ID NO: 433 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 427.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 426; the variable heavy chain region of SEQID NO: 427; the complementarity-determining regions (SEQ ID NO: 428; SEQID NO: 429; and SEQ ID NO: 430) of the variable light chain region ofSEQ ID NO: 426; and the complementarity-determining regions (SEQ ID NO:431; SEQ ID NO: 432; and SEQ ID NO: 433) of the variable heavy chainregion of SEQ ID NO: 427.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb27, comprising SEQ ID NO: 426 and SEQ ID NO: 427, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 442) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSEPVGGTVSISCQSSKSVMNNNYLAWYQQKPGQPPKLLIYGASNLASGVPSRFSGSGSGTQFTLTISDVQCDDAATYYCQGGYTGYSDHGT

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 443) METGLRWLLLVAVLKGVQCQSVEESGGRLVKPDETLTLTCTVSGIDLSSYPMNWVRQAPGKGLEWIGFINTGGTIVYASWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARGSYVSSGYAYYFNV.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 444; SEQ ID NO: 445; and SEQ IDNO: 446 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 442, and/or one or more of the polypeptide sequences of SEQID NO: 447; SEQ ID NO: 448; and SEQ ID NO: 449 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 443, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 444; SEQ ID NO: 445; and SEQ ID NO:446 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 442, and/or one or more of the polypeptide sequences of SEQ ID NO:447; SEQ ID NO: 448; and SEQ ID NO: 449 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 443, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 442. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 443.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 444; SEQ IDNO: 445; and SEQ ID NO: 446 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 442.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 447; SEQ IDNO: 448; and SEQ ID NO: 449 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 443.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 442; the variable heavy chain region of SEQID NO: 443; the complementarity-determining regions (SEQ ID NO: 444; SEQID NO: 445; and SEQ ID NO: 446) of the variable light chain region ofSEQ ID NO: 442; and the complementarity-determining regions (SEQ ID NO:447; SEQ ID NO: 448; and SEQ ID NO: 449) of the variable heavy chainregion of SEQ ID NO: 443.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb28, comprising SEQ ID NO: 442 and SEQ ID NO: 443, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 458) MDTRAPTQLLGLLLLWLPGATFAAVLTQTPSPVSAAVGGTVSISCQSSQSVYNNNWLSWFQQKPGQPPKLLIYKASTLASGVPSRFKGSGSGTQFTLTIS DVQCDDVATYYCAGGYLDSVI

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 459) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSTYSINWVRQAPGKGLEWIGIIANSGTTFYANWAKGRFTVSKTSTTVDLKITSPTTEDTATYFCARESGMYNEYGKFNI.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 460; SEQ ID NO: 461; and SEQ IDNO: 462 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 458, and/or one or more of the polypeptide sequences of SEQID NO: 463; SEQ ID NO: 464; and SEQ ID NO: 465 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 459, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 460; SEQ ID NO: 461; and SEQ ID NO:462 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 458, and/or one or more of the polypeptide sequences of SEQ ID NO:463; SEQ ID NO: 464; and SEQ ID NO: 465 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 459, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 458. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 459.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 460; SEQ IDNO: 461; and SEQ ID NO: 462 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 458.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 463; SEQ IDNO: 464; and SEQ ID NO: 465 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 459.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 458; the variable heavy chain region of SEQID NO: 459; the complementarity-determining regions (SEQ ID NO: 460; SEQID NO: 461; and SEQ ID NO: 462) of the variable light chain region ofSEQ ID NO: 458; and the complementarity-determining regions (SEQ ID NO:463; SEQ ID NO: 464; and SEQ ID NO: 465) of the variable heavy chainregion of SEQ ID NO: 459.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb29, comprising SEQ ID NO: 458 and SEQ ID NO: 459, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 474) MDTRAPTQLLGLLLLWLPGARCASDMTQTPSSVSAAVGGTVTINCQASENIYSFLAWYQQKPGQPPKLLIFKASTLASGVSSRFKGSGSGTQFTLTISDLECDDAATYYCQQGATVYDIDNN

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 475) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGIDLSAYAMIWVRQAPGEGLEWITITYPNGITYYANWAKGRFTVSKTSTAMDLKITSPTTEDTATYFCARDAESSKNAYWGYFNV.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 476; SEQ ID NO: 477; and SEQ IDNO: 478 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 474, and/or one or more of the polypeptide sequences of SEQID NO: 479; SEQ ID NO: 480; and SEQ ID NO: 481 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 475, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 476; SEQ ID NO: 477; and SEQ ID NO:478 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 474, and/or one or more of the polypeptide sequences of SEQ ID NO:479; SEQ ID NO: 480; and SEQ ID NO: 481 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 475, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 474. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 475.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 476; SEQ IDNO: 477; and SEQ ID NO: 478 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 474.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 479; SEQ IDNO: 480; and SEQ ID NO: 481 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 475.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 474; the variable heavy chain region of SEQID NO: 475; the complementarity-determining regions (SEQ ID NO: 476; SEQID NO: 477; and SEQ ID NO: 478) of the variable light chain region ofSEQ ID NO: 474; and the complementarity-determining regions (SEQ ID NO:479; SEQ ID NO: 480; and SEQ ID NO: 481) of the variable heavy chainregion of SEQ ID NO: 475.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb30, comprising SEQ ID NO: 474 and SEQ ID NO: 475, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 490) MDTRAPTQLLGLLLLWLPGARCASDMTQTPSSVSAAVGGTVTINCQASENIYSFLAWYQQKPGQPPKLLIFRASTLASGVSSRFKGSGSGTQFTLTISDLECDDAATYYCQQGATVYDIDNN

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 491) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGIDLSAYAMIWVRQAPGEGLEWITIIYPNGITYYANWAKGRFTVSKTSTAMDLKITSPTTEDTATYFCARDAESSKNAYWGYFNV.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 492; SEQ ID NO: 493; and SEQ IDNO: 494 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 490, and/or one or more of the polypeptide sequences of SEQID NO: 495; SEQ ID NO: 496; and SEQ ID NO: 497 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 491, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 492; SEQ ID NO: 493; and SEQ ID NO:494 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 490, and/or one or more of the polypeptide sequences of SEQ ID NO:495; SEQ ID NO: 496; and SEQ ID NO: 497 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 491, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 490. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 491.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 492; SEQ IDNO: 493; and SEQ ID NO: 494 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 490.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 495; SEQ IDNO: 496; and SEQ ID NO: 497 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 491.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 490; the variable heavy chain region of SEQID NO: 491; the complementarity-determining regions (SEQ ID NO: 492; SEQID NO: 493; and SEQ ID NO: 494) of the variable light chain region ofSEQ ID NO: 490; and the complementarity-determining regions (SEQ ID NO:495; SEQ ID NO: 496; and SEQ ID NO: 497) of the variable heavy chainregion of SEQ ID NO: 491.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb31, comprising SEQ ID NO: 490 and SEQ ID NO: 491, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 506) MDTRAPTQLLGLLLLWLPGATFAIEMTQTPSPVSAAVGGTVTINCQASESVFNNMLSWYQQKPGHSPKLLIYDASDLASGVPSRFKGSGSGTQFTLTISGVECDDAATYYCAGYKSDSNDGDNV

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 507) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLNRNSITWVRQAPGEGLEWIGIITGSGRTYYANWAKGRFTISKTSTTVDLKMTSPTTEDTATYFCARGHPGLGSGNI.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 508; SEQ ID NO: 509; and SEQ IDNO: 510 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 506, and/or one or more of the polypeptide sequences of SEQID NO: 511; SEQ ID NO: 512; and SEQ ID NO: 513 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 507, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 508; SEQ ID NO: 509; and SEQ ID NO:510 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 506, and/or one or more of the polypeptide sequences of SEQ ID NO:511; SEQ ID NO: 512; and SEQ ID NO: 513 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 507, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 506. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 507.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 508; SEQ IDNO: 509; and SEQ ID NO: 510 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 506.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 511; SEQ IDNO: 512; and SEQ ID NO: 513 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 507.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 506; the variable heavy chain region of SEQID NO: 507; the complementarity-determining regions (SEQ ID NO: 508; SEQID NO: 509; and SEQ ID NO: 510) of the variable light chain region ofSEQ ID NO: 506; and the complementarity-determining regions (SEQ ID NO:511; SEQ ID NO: 512; and SEQ ID NO: 513) of the variable heavy chainregion of SEQ ID NO: 507.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb32, comprising SEQ ID NO: 506 and SEQ ID NO: 507, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 522) MDTRAPTQLLGLLLLWLPGATFAQVLTQTASSVSAAVGGTVTINCQSSQSVYNNYLSWYQQKPGQPPKLLIYTASSLASGVPSRFKGSGSGTQFTLTISE VQCDDAATYYCQGYYSGPIIT

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 523) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTASGFSLNNYYIQWVRQAPGEGLEWIGIIYAGGSAYYATWANGRFTIAKTSSTTVDLKMTSLTTEDTATYFCARGTFDGYEL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 524; SEQ ID NO: 525; and SEQ IDNO: 526 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 522, and/or one or more of the polypeptide sequences of SEQID NO: 527; SEQ ID NO: 528; and SEQ ID NO: 529 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 523, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 524; SEQ ID NO: 525; and SEQ ID NO:526 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 522, and/or one or more of the polypeptide sequences of SEQ ID NO:527; SEQ ID NO: 528; and SEQ ID NO: 529 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 523, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 522. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 523.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 524; SEQ IDNO: 525; and SEQ ID NO: 526 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 522.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 527; SEQ IDNO: 528; and SEQ ID NO: 529 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 523.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 522; the variable heavy chain region of SEQID NO: 523; the complementarity-determining regions (SEQ ID NO: 524; SEQID NO: 525; and SEQ ID NO: 526) of the variable light chain region ofSEQ ID NO: 522; and the complementarity-determining regions (SEQ ID NO:527; SEQ ID NO: 528; and SEQ ID NO: 529) of the variable heavy chainregion of SEQ ID NO: 523.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb33, comprising SEQ ID NO: 522 and SEQ ID NO: 523, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 538) MDTRAPTQLLGLLLLWLPGATFAQVLTQTPSPVSVPVGDTVTISCQSSESVYSNNLLSWYQQKPGQPPKLLIYRASNLASGVPSRFKGSGSGTQFTLTISGAQCDDAATYYCQGYYSGVINS

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 539) METGLRWLLLVAVLKGVQCQSVEESGGRLVTPGTPLTLTCTVSGFSLSSYFMSWVRQAPGEGLEYIGFINPGGSAYYASWASGRLTISKTSTTVDLKITSPTTEDTATYFCARILIVSYGAFTI.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 540; SEQ ID NO: 541; and SEQ IDNO: 542 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 538, and/or one or more of the polypeptide sequences of SEQID NO: 543; SEQ ID NO: 544; and SEQ ID NO: 545 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 539, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 540; SEQ ID NO: 541; and SEQ ID NO:542 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 538, and/or one or more of the polypeptide sequences of SEQ ID NO:543; SEQ ID NO: 544; and SEQ ID NO: 545 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 539, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 538. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 539.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 540; SEQ IDNO: 541; and SEQ ID NO: 542 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 538.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 543; SEQ IDNO: 544; and SEQ ID NO: 545 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 539.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 538; the variable heavy chain region of SEQID NO: 539; the complementarity-determining regions (SEQ ID NO: 540; SEQID NO: 541; and SEQ ID NO: 542) of the variable light chain region ofSEQ ID NO: 538; and the complementarity-determining regions (SEQ ID NO:543; SEQ ID NO: 544; and SEQ ID NO: 545) of the variable heavy chainregion of SEQ ID NO: 539.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb34, comprising SEQ ID NO: 538 and SEQ ID NO: 539, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 554) MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQATESIGNELSWYQQKPGQAPKLLIYSASTLASGVPSRFKGSGSGTQFTLTITGVECDDAATYYCQQGYSSANIDNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 555) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSKYYMSWVRQAPEKGLKYIGYIDSTTVNTYYATWARGRFTISKTSTTVDLKITSPTSEDTATYFCARGSTYFTDGGHRLDL.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 556; SEQ ID NO: 557; and SEQ IDNO: 558 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 554, and/or one or more of the polypeptide sequences of SEQID NO: 559; SEQ ID NO: 560; and SEQ ID NO: 561 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 555, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 556; SEQ ID NO: 557; and SEQ ID NO:558 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 554, and/or one or more of the polypeptide sequences of SEQ ID NO:559; SEQ ID NO: 560; and SEQ ID NO: 561 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 555, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 554. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 555.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 556; SEQ IDNO: 557; and SEQ ID NO: 558 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 554.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 559; SEQ IDNO: 560; and SEQ ID NO: 561 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 555.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 554; the variable heavy chain region of SEQID NO: 555; the complementarity-determining regions (SEQ ID NO: 556; SEQID NO: 557; and SEQ ID NO: 558) of the variable light chain region ofSEQ ID NO: 554; and the complementarity-determining regions (SEQ ID NO:559; SEQ ID NO: 560; and SEQ ID NO: 561) of the variable heavy chainregion of SEQ ID NO: 555.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb35, comprising SEQ ID NO: 554 and SEQ ID NO: 555, and having at leastone of the biological activities set forth herein.

In another embodiment, the invention includes antibodies having bindingspecificity to IL-6 and possessing a variable light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 570) MDTRAPTQLLGLLLLWLPGARCAYDMTQTPASVEVAVGGTVTIKCQATESIGNELSWYQQKPGQAPKLLIYSASTLASGVPSRFKGSGSGTQFTLTITGVECDDAATYYCQQGYSSANIDNA

The invention also includes antibodies having binding specificity toIL-6 and possessing a variable heavy chain sequence comprising thesequence set forth below:

(SEQ ID NO: 571) METGLRWLLLVAVLKGVQCQSLEESGGRLVTPGTPLTLTCTVSGFSLSTYNMGWVRQAPGKGLEWIGSITIDGRTYYASWAKGRFTVSKSSTTVDLKMTSLTTGDTATYFCARILIVSYGAFTI.

The invention further contemplates antibodies comprising one or more ofthe polypeptide sequences of SEQ ID NO: 572; SEQ ID NO: 573; and SEQ IDNO: 574 which correspond to the complementarity-determining regions(CDRs, or hypervariable regions) of the variable light chain sequence ofSEQ ID NO: 570, and/or one or more of the polypeptide sequences of SEQID NO: 575; SEQ ID NO: 576; and SEQ ID NO: 577 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 571, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

In another embodiment, the invention contemplates other antibodies, suchas for example chimeric antibodies, comprising one or more of thepolypeptide sequences of SEQ ID NO: 572; SEQ ID NO: 573; and SEQ ID NO:574 which correspond to the complementarity-determining regions (CDRs,or hypervariable regions) of the variable light chain sequence of SEQ IDNO: 570, and/or one or more of the polypeptide sequences of SEQ ID NO:575; SEQ ID NO: 576; and SEQ ID NO: 577 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 571, or combinations ofthese polypeptide sequences. In another embodiment of the invention, theantibodies of the invention include combinations of the CDRs and thevariable heavy and light chain sequences set forth above.

The invention also contemplates fragments of the antibody having bindingspecificity to IL-6. In one embodiment of the invention, antibodyfragments of the invention comprise, or alternatively consist of, thepolypeptide sequence of SEQ ID NO: 570. In another embodiment of theinvention, antibody fragments of the invention comprise, oralternatively consist of, the polypeptide sequence of SEQ ID NO: 571.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 572; SEQ IDNO: 573; and SEQ ID NO: 574 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable light chain sequence of SEQ ID NO: 570.

In a further embodiment of the invention, fragments of the antibodyhaving binding specificity to IL-6 comprise, or alternatively consistof, one or more of the polypeptide sequences of SEQ ID NO: 575; SEQ IDNO: 576; and SEQ ID NO: 577 which correspond to thecomplementarity-determining regions (CDRs, or hypervariable regions) ofthe variable heavy chain sequence of SEQ ID NO: 571.

The invention also contemplates antibody fragments which include one ormore of the antibody fragments described herein. In one embodiment ofthe invention, fragments of the antibodies having binding specificity toIL-6 comprise, or alternatively consist of, one, two, three or more,including all of the following antibody fragments: the variable lightchain region of SEQ ID NO: 570; the variable heavy chain region of SEQID NO: 571; the complementarity-determining regions (SEQ ID NO: 572; SEQID NO: 573; and SEQ ID NO: 574) of the variable light chain region ofSEQ ID NO: 570; and the complementarity-determining regions (SEQ ID NO:575; SEQ ID NO: 576; and SEQ ID NO: 577) of the variable heavy chainregion of SEQ ID NO: 571.

In a preferred embodiment of the invention, the anti-IL-6 antibody isAb36, comprising SEQ ID NO: 570 and SEQ ID NO: 571, and having at leastone of the biological activities set forth herein.

Such antibody fragments may be present in one or more of the followingnon-limiting forms: Fab, Fab′, F(ab′)₂, Fv and single chain Fv antibodyforms. In a preferred embodiment, the anti-IL-6 antibodies describedherein further comprises the kappa constant light chain sequencecomprising the sequence set forth below:

(SEQ ID NO: 586) VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSF NRGEC.

In another preferred embodiment, the anti-IL-6 antibodies describedherein further comprises and the gamma-1 constant heavy chainpolypeptide sequence comprising the sequence set forth below:

(SEQ ID NO: 588) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK.

In another embodiment, the invention contemplates an isolated anti-IL-6antibody comprising a V_(H) polypeptide sequence selected from the groupconsisting of: SEQ ID NO: 3, 18, 19, 22, 38, 54, 70, 86, 102, 117, 118,123, 139, 155, 171, 187, 203, 219, 235, 251, 267, 283, 299, 315, 331,347, 363, 379, 395, 411, 427, 443, 459, 475, 491, 507, 523, 539, 555 andSEQ ID NO: 571; and further comprising a V_(L) polypeptide sequenceselected from the group consisting of: SEQ ID NO: 2, 20, 21, 37, 53, 69,85, 101, 119, 122, 138, 154, 170, 186, 202, 218, 234, 250, 266, 282,298, 314, 330, 346, 362, 378, 394, 410, 426, 442, 458, 474, 490, 506,522, 538, 554 and SEQ ID NO: 570 or a variant thereof wherein one ormore of the framework residues (FR residues) in said V_(H) or V_(L)polypeptide has been substituted with another amino acid residueresulting in an anti-IL-6 antibody that specifically binds IL-6. Theinvention contemplates humanized and chimeric forms of these antibodies.The chimeric antibodies may include an Fc derived from IgG1, IgG2, IgG3,IgG4, IgG5, IgG6, IgG7, IgG8, IgG9, IgG10, IgG11, IgG12, IgG13, IgG14,IgG15, IgG16, IgG17, IgG18 or IgG19 constant regions.

In one embodiment of the invention, the antibodies or V_(H) or V_(L)polypeptides originate or are selected from one or more rabbit B cellpopulations prior to initiation of the humanization process referencedherein.

In another embodiment of the invention, the anti-IL-6 antibodies andfragments thereof have binding specificity for primate homologs of thehuman IL-6 protein. Non-limiting examples of primate homologs of thehuman IL-6 protein are IL-6 obtained from Macacaca fascicularis (alsoknown as the cynomolgus monkey) and the Rhesus monkey. In anotherembodiment of the invention, the anti-IL-6 antibodies and fragmentsthereof inhibits the association of IL-6 with IL-6R, and/or theproduction of IL-6/IL-6R/gp30 complexes and/or the production ofIL-6/IL-6R/gp30 multimers and/or antagonizes the biological effects ofone or more of the foregoing.

As stated above, antibodies and fragments thereof may be modifiedpost-translationally to add effector moieties such as chemical linkers,detectable moieties such as for example fluorescent dyes, enzymes,substrates, bioluminescent materials, radioactive materials, andchemiluminescent moieties, or functional moieties such as for examplestreptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, andradioactive materials.

Regarding detectable moieties, further exemplary enzymes include, butare not limited to, horseradish peroxidase, acetylcholinesterase,alkaline phosphatase, beta-galactosidase and luciferase. Furtherexemplary fluorescent materials include, but are not limited to,rhodamine, fluorescein, fluorescein isothiocyanate, umbelliferone,dichlorotriazinylamine, phycoerythrin and dansyl chloride. Furtherexemplary chemiluminescent moieties include, but are not limited to,luminol. Further exemplary bioluminescent materials include, but are notlimited to, luciferin and aequorin. Further exemplary radioactivematerials include, but are not limited to, Iodine 125 (¹²⁵I), Carbon 14(¹⁴C), Sulfur 35 (³⁵S), Tritium (³H) and Phosphorus 32 (³²P).

Regarding functional moieties, exemplary cytotoxic agents include, butare not limited to, methotrexate, aminopterin, 6-mercaptopurine,6-thioguanine, cytarabine, 5-fluorouracil decarbazine; alkylating agentssuch as mechlorethamine, thioepa chlorambucil, melphalan, carmustine(BSNU), mitomycin C, lomustine (CCNU), I-methylnitrosourea,cyclothosphamide, mechlorethamine, busulfan, dibromomannitol,streptozotocin, mitomycin C, cis-dichlorodiamine platinum (II) (DDP)cisplatin and carboplatin (paraplatin); anthracyclines includedaunorubicin (formerly daunomycin), doxorubicin (adriamycin),detorubicin, carminomycin, idarubicin, epirubicin, mitoxantrone andbisantrene; antibiotics include dactinomycin (actinomycin D), bleomycin,calicheamicin, mithramycin, and anthramycin (AMC); and antimytoticagents such as the vinca alkaloids, vincristine and vinblastine. Othercytotoxic agents include paclitaxel (taxol), ricin, pseudomonasexotoxin, gemcitabine, cytochalasin B, gramicidin D, ethidium bromide,emetine, etoposide, tenoposide, colchicin, dihydroxy anthracin dione,1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine,propranolol, puromycin, procarbazine, hydroxyurea, asparaginase,corticosteroids, mytotane (O,P′-(DDD)), interferons, and mixtures ofthese cytotoxic agents.

Further cytotoxic agents include, but are not limited to,chemotherapeutic agents such as carboplatin, cisplatin, paclitaxel,gemcitabine, calicheamicin, doxorubicin, 5-fluorouracil, mitomycin C,actinomycin D, cyclophosphamide, vincristine, bleomycin, VEGFantagonists, EGFR antagonists, platins, taxols, irinotecan,5-fluorouracil, gemcytabine, leucovorine, steroids, cyclophosphamide,melphalan, vinca alkaloids (e.g., vinblastine, vincristine, vindesineand vinorelbine), mustines, tyrosine kinase inhibitors, radiotherapy,sex hormone antagonists, selective androgen receptor modulators,selective estrogen receptor modulators, PDGF antagonists, TNFantagonists, IL-1 antagonists, interleukins (e.g. IL-12 or IL-2), IL-12Rantagonists, Toxin conjugated monoclonal antibodies, tumor antigenspecific monoclonal antibodies, Erbitux, Avastin, Pertuzumab, anti-CD20antibodies, Rituxan, ocrelizumab, ofatumumab, DXL625, herceptin, or anycombination thereof. Toxic enzymes from plants and bacteria such asricin, diphtheria toxin and Pseudomonas toxin may be conjugated to thehumanized antibodies, or binding fragments thereof, to generatecell-type-specific-killing reagents (Youle, et al., Proc. Nat'l Acad.Sci. USA 77:5483 (1980); Gilliland, et al., Proc. Nat'l Acad. Sci. USA77:4539 (1980); Krolick, et al., Proc. Nat'l Acad. Sci. USA 77:5419(1980)).

Other cytotoxic agents include cytotoxic ribonucleases as described byGoldenberg in U.S. Pat. No. 6,653,104. Embodiments of the invention alsorelate to radioimmunoconjugates where a radionuclide that emits alpha orbeta particles is stably coupled to the antibody, or binding fragmentsthereof, with or without the use of a complex-forming agent. Suchradionuclides include beta-emitters such as Phosphorus-32 (³²P),Scandium-47 (⁴⁷Sc), Copper-67 (⁶⁷Cu), Gallium-67 (⁶⁷Ga), Yttrium-88(⁸⁸Y), Yttrium-90 (⁹⁰Y), Iodine-125 (¹²⁵I), Iodine-131 (¹³¹I),Samarium-153 (¹⁵³Sm), Lutetium-177 (¹⁷⁷Lu), Rhenium-186 (¹⁸⁶Re) orRhenium-188 (¹⁸Re), and alpha-emitters such as Astatine-211 (²¹¹At),Lead-212 (²¹²Pb), Bismuth-212 (²¹²Bi) or -213 (²¹³Bi) or Actinium-225(²²⁵Ac).

Methods are known in the art for conjugating an antibody or bindingfragment thereof to a detectable moiety and the like, such as forexample those methods described by Hunter et al, Nature 144:945 (1962);David et al, Biochemistry 13:1014 (1974); Pain et al, J. Immunol. Meth.40:219 (1981); and Nygren, J., Histochem. and Cytochem. 30:407 (1982).

Embodiments described herein further include variants and equivalentsthat are substantially homologous to the antibodies, antibody fragments,diabodies, SMIPs, camelbodies, nanobodies, IgNAR, polypeptides, variableregions and CDRs set forth herein. These may contain, e.g., conservativesubstitution mutations, (i.e., the substitution of one or more aminoacids by similar amino acids). For example, conservative substitutionrefers to the substitution of an amino acid with another within the samegeneral class, e.g., one acidic amino acid with another acidic aminoacid, one basic amino acid with another basic amino acid, or one neutralamino acid by another neutral amino acid. What is intended by aconservative amino acid substitution is well known in the art.

In another embodiment, the invention contemplates polypeptide sequenceshaving at least 90% or greater sequence homology to any one or more ofthe polypeptide sequences of antibody fragments, variable regions andCDRs set forth herein. More preferably, the invention contemplatespolypeptide sequences having at least 95% or greater sequence homology,even more preferably at least 98% or greater sequence homology, andstill more preferably at least 99% or greater sequence homology to anyone or more of the polypeptide sequences of antibody fragments, variableregions and CDRs set forth herein. Methods for determining homologybetween nucleic acid and amino acid sequences are well known to those ofordinary skill in the art.

In another embodiment, the invention further contemplates theabove-recited polypeptide homologs of the antibody fragments, variableregions and CDRs set forth herein further having anti-IL-6 activity.Non-limiting examples of anti-IL-6 activity are set forth herein, forexample, under the heading “Anti-IL-6 Activity,” infra.

In another embodiment, the invention further contemplates the generationand use of anti-idiotypic antibodies that bind any of the foregoingsequences. In an exemplary embodiment, such an anti-idiotypic antibodycould be administered to a subject who has received an anti-IL-6antibody to modulate, reduce, or neutralize, the effect of the anti-IL-6antibody. Such anti-idiotypic antibodies could also be useful fortreatment of an autoimmune disease characterized by the presence ofanti-IL-6 antibodies. A further exemplary use of such anti-idiotypicantibodies is for detection of the anti-IL-6 antibodies of the presentinvention, for example to monitor the levels of the anti-IL-6 antibodiespresent in a subject's blood or other bodily fluids.

The present invention also contemplates anti-IL-6 antibodies comprisingany of the polypeptide or polynucleotide sequences described hereinsubstituted for any of the other polynucleotide sequences describedherein. For example, without limitation thereto, the present inventioncontemplates antibodies comprising the combination of any of thevariable light chain and variable heavy chain sequences describedherein, and further contemplates antibodies resulting from substitutionof any of the CDR sequences described herein for any of the other CDRsequences described herein.

Additional Exemplary Embodiments of the Invention

In another embodiment, the invention contemplates one or more anti-IL-6antibodies or antibody fragment which may specifically bind to the samelinear or conformational epitope(s) and/or compete for binding to thesame linear or conformational epitope(s) on an intact human IL-6polypeptide or fragment thereof as an anti-IL-6 antibody selected fromthe group consisting of Ab1, Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9,Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21,Ab22, Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33,Ab34, Ab35, and Ab36. In a preferred embodiment, the anti-IL-6 antibodyor fragment may specifically bind to the same linear or conformationalepitope(s) and/or compete for binding to the same linear orconformational epitope(s) on an intact human IL-6 polypeptide or afragment thereof as Ab1.

In another embodiment of the invention, the anti-IL-6 antibody whichspecifically binds to the same linear or conformational epitopes on anintact IL-6 polypeptide or fragment thereof that is (are) specificallybound by Ab1 binds to a IL-6 epitope(s) ascertained by epitopic mappingusing overlapping linear peptide fragments which span the full length ofthe native human IL-6 polypeptide. In one embodiment of the invention,the IL-6 epitope comprises, or alternatively consists of, one or moreresidues comprised in IL-6 fragments selected from those respectivelyencompassing amino acid residues 37-51, amino acid residues 70-84, aminoacid residues 169-183, amino acid residues 31-45 and/or amino acidresidues 58-72.

The invention is also directed to an anti-IL-6 antibody that binds withthe same IL-6 epitope and/or competes with an anti-IL-6 antibody forbinding to IL-6 as an antibody or antibody fragment disclosed herein,including but not limited to an anti-IL-6 antibody selected from Ab1,Ab2, Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14,Ab15, Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25, Ab26,Ab27, Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, and Ab36.

In another embodiment, the invention is also directed to an isolatedanti-IL-6 antibody or antibody fragment comprising one or more of theCDRs contained in the V_(H) polypeptide sequences selected from thegroup consisting of: SEQ ID NO: 3, 18, 19, 22, 38, 54, 70, 86, 102, 117,118, 123, 139, 155, 171, 187, 203, 219, 235, 251, 267, 283, 299, 315,331, 347, 363, 379, 395, 411, 427, 443, 459, 475, 491, 507, 523, 539,555 and SEQ ID NO: 571 and/or one or more of the CDRs contained in theV_(L) polypeptide sequence consisting of: 2, 20, 21, 37, 53, 69, 85,101, 119, 122, 138, 154, 170, 186, 202, 218, 234, 250, 266, 282, 298,314, 330, 346, 362, 378, 394, 410, 426, 442, 458, 474, 490, 506, 522,538, 554 and SEQ ID NO: 570.

In one embodiment of the invention, the anti-IL-6 antibody discussed inthe two prior paragraphs comprises at least 2 complementaritydetermining regions (CDRs) in each the variable light and the variableheavy regions which are identical to those contained in an anti-IL-6antibody selected from the group consisting of Ab1, Ab2, Ab3, Ab4, Ab5,Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15, Ab16, Ab17,Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25, Ab26, Ab27, Ab28, Ab29,Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, and Ab36.

In a preferred embodiment, the anti-IL-6 antibody discussed abovecomprises at least 2 complementarity determining regions (CDRs) in eachthe variable light and the variable heavy regions which are identical tothose contained in Ab1. In another embodiment, all of the CDRs of theanti-IL-6 antibody discussed above are identical to the CDRs containedin an anti-IL-6 antibody selected from the group consisting of Ab1, Ab2,Ab3, Ab4, Ab5, Ab6, Ab7, Ab8, Ab9, Ab10, Ab11, Ab12, Ab13, Ab14, Ab15,Ab16, Ab17, Ab18, Ab19, Ab20, Ab21, Ab22, Ab23, Ab24, Ab25, Ab26, Ab27,Ab28, Ab29, Ab30, Ab31, Ab32, Ab33, Ab34, Ab35, and Ab36. In a preferredembodiment of the invention, all of the CDRs of the anti-IL-6 antibodydiscussed above are identical to the CDRs contained in Ab1.

The invention further contemplates that the one or more anti-IL-6antibodies discussed above are aglycosylated; that contain an Fc regionthat has been modified to alter effector function, half-life,proteolysis, and/or glycosylation; are human, humanized, single chain orchimeric; and are a humanized antibody derived from a rabbit (parent)anti-IL-6 antibody.

The invention further contemplates one or more anti-IL-6 antibodieswherein the framework regions (FRs) in the variable light region and thevariable heavy regions of said antibody respectively are human FRs whichare unmodified or which have been modified by the substitution of atmost 2 or 3 human FR residues in the variable light or heavy chainregion with the corresponding FR residues of the parent rabbit antibody,and wherein said human FRs have been derived from human variable heavyand light chain antibody sequences which have been selected from alibrary of human germline antibody sequences based on their high levelof homology to the corresponding rabbit variable heavy or light chainregions relative to other human germline antibody sequences contained inthe library.

In one embodiment of the invention, the anti-IL-6 antibody or fragmentmay specifically bind to IL-6 expressing human cells and/or tocirculating soluble IL-6 molecules in vivo, including IL-6 expressed onor by human cells in a patient with a disease associated with cells thatexpress IL-6.

In another embodiment, the disease is selected from general fatigue,exercise-induced fatigue, cancer-related fatigue, inflammatorydisease-related fatigue, chronic fatigue syndrome, fibromyalgia,cancer-related cachexia, cardiac-related cachexia, respiratory-relatedcachexia, renal-related cachexia, age-related cachexia, rheumatoidarthritis, systemic lupus erythematosis (SLE), systemic juvenileidiopathic arthritis, psoriasis, psoriatic arthropathy, ankylosingspondylitis, inflammatory bowel disease (IBD), polymyalgia rheumatica,giant cell arteritis, autoimmune vasculitis, graft versus host disease(GVHD), Sjogren's syndrome, adult onset Still's disease, rheumatoidarthritis, systemic juvenile idiopathic arthritis, osteoarthritis,osteoporosis, Paget's disease of bone, osteoarthritis, multiple myeloma,Hodgkin's lymphoma, non-Hodgkin's lymphoma, prostate cancer, leukemia,renal cell cancer, multicentric Castleman's disease, ovarian cancer,drug resistance in cancer chemotherapy, cancer chemotherapy toxicity,ischemic heart disease, atherosclerosis, obesity, diabetes, asthma,multiple sclerosis, Alzheimer's disease, cerebrovascular disease, fever,acute phase response, allergies, anemia, anemia of inflammation (anemiaof chronic disease), hypertension, depression, depression associatedwith a chronic illness, thrombosis, thrombocytosis, acute heart failure,metabolic syndrome, miscarriage, obesity, chronic prostatitis,glomerulonephritis, pelvic inflammatory disease, reperfusion injury,transplant rejection, graft versus host disease (GVHD), avian influenza,smallpox, pandemic influenza, adult respiratory distress syndrome(ARDS), severe acute respiratory syndrome (SARS), sepsis, and systemicinflammatory response syndrome (SIRS). In a preferred embodiment, thedisease is selected from a cancer, inflammatory disorder, viraldisorder, or autoimmune disorder. In a particularly preferredembodiment, the disease is arthritis, cachexia, and wasting syndrome

The invention further contemplates anti-IL-6 antibodies or fragmentsdirectly or indirectly attached to a detectable label or therapeuticagent.

The invention also contemplates one or more nucleic acid sequences whichresult in the expression of an anti-IL-6 antibody or antibody fragmentas set forth above, including those comprising, or alternativelyconsisting of, yeast or human preferred codons. The invention alsocontemplates vectors (including plasmids or recombinant viral vectors)comprising said nucleic acid sequence(s). The invention alsocontemplates host cells or recombinant host cells expressing at leastone of the antibodies set forth above, including a mammalian, yeast,bacterial, and insect cells. In a preferred embodiment, the host cell isa yeast cell. In a further preferred embodiment, the yeast cell is adiploidal yeast cell. In a more preferred embodiment, the yeast cell isa Pichia yeast.

The invention also contemplates a method of treatment comprisingadministering to a patient with a disease or condition associated withIL-6 expressing cells a therapeutically effective amount of at least oneanti-IL-6 antibody or fragment. The diseases that may be treated arepresented in the non-limiting list set forth above. In a preferredembodiment, the disease is selected from a cancer, autoimmune disease,or inflammatory condition. In a particularly preferred embodiment, thedisease is cancer or viral infection. In another embodiment thetreatment further includes the administration of another therapeuticagent or regimen selected from chemotherapy, radiotherapy, cytokineadministration or gene therapy.

The invention further contemplates a method of in vivo imaging whichdetects the presence of cells which express IL-6 comprisingadministering a diagnostically effective amount of at least oneanti-IL-6 antibody. In one embodiment, said administration furtherincludes the administration of a radionuclide or fluorophore thatfacilitates detection of the antibody at IL-6 expressing disease sites.In another embodiment of the invention, the method of in vivo imaging isused to detect IL-6 expressing tumors or metastases or is used to detectthe presence of sites of autoimmune disorders associated with IL-6expressing cells. In a further embodiment, the results of said in vivoimaging method are used to facilitate design of an appropriatetherapeutic regimen, including therapeutic regimens includingradiotherapy, chemotherapy or a combination thereof.

Polynucleotides Encoding Anti-IL-6 Antibody Polypeptides

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 2:

(SEQ ID NO: 10) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAACCAGGGCAGCGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 3:

(SEQ ID NO: 11) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTAACTACTACGTGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAATCATTTATGGTAGTGATGAAACGGCCTACGCGACCTGGGCGATAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAACTTGTGGGGCCAAGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 12; SEQ ID NO: 13; and SEQ ID NO: 14 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 2.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 15; SEQ ID NO: 16; and SEQ ID NO: 17 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 3.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 10 encoding the light chainvariable region of SEQ ID NO: 2; the polynucleotide SEQ ID NO: 11encoding the heavy chain variable region of SEQ ID NO: 3;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 12; SEQ ID NO: 13; and SEQ ID NO: 14) of the light chain variableregion of SEQ ID NO: 10; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 15; SEQ ID NO: 16; andSEQ ID NO: 17) of the heavy chain variable region of SEQ ID NO: 11.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 21:

(SEQ ID NO: 29) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTGAGACCATTTACAGTTGGTTATCCTGGTATCAGCAGAAGCCAGGGCAGCCTCCCAAGCTCCTGATCTACCAGGCATCCGATCTGGCATCTGGGGTCCCATCGCGATTCAGCGGCAGTGGGGCTGGGACAGAGTACACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCAACAGGGTTATAGTGGTAGTAATGTTGATAATGTTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAG AGAGGCCAAAG

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 22:

(SEQ ID NO: 30) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGGAGCAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTTACCTGCACAGCCTCTGGATTCTCCCTCAATGACCATGCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGATTCATTAATAGTGGTGGTAGCGCACGCTACGCGAGCTGGGCAGAAGGCCGATTCACCATCTCCAGAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTGTCAGAGGGGGTGCTGTTTGGAGTATTCATAGTTTTGATCCCTGGGGCCCAGGGACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAA G.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 31; SEQ ID NO: 32; and SEQ ID NO: 33 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 21.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 34; SEQ ID NO: 35; and SEQ ID NO: 36 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 22.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 29 encoding the light chainvariable region of SEQ ID NO: 21; the polynucleotide SEQ ID NO: 30encoding the heavy chain variable region of SEQ ID NO: 22;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 3I; SEQ ID NO: 32; and SEQ ID NO: 33) of the light chain variableregion of SEQ ID NO: 29; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 34; SEQ ID NO: 35; andSEQ ID NO: 36) of the heavy chain variable region of SEQ ID NO: 30.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 37:

(SEQ ID NO: 45) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGGCCAGTCAGAGTGTTTATGACAACAACTACTTATCCTGGTTTCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCGTGGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACAGACGTGCAGTGTGACGATGCTGCCACTTACTATTGTGCAGGCGTTTATGATGATGATAGTGATAATGCCTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 38:

(SEQ ID NO: 46) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTGGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACCCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTGTCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGATTCATTACAATGAGTGATAATATAAATTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTACAATGGGTCGGTTGGATCTCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 47; SEQ ID NO: 48; and SEQ ID NO: 49 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 37.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 50; SEQ ID NO: 51; and SEQ ID NO: 52 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 38.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 45 encoding the light chainvariable region of SEQ ID NO: 37; the polynucleotide SEQ ID NO: 46encoding the heavy chain variable region of SEQ ID NO: 38;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 47; SEQ ID NO: 48; and SEQ ID NO: 49) of the light chain variableregion of SEQ ID NO: 37; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 50; SEQ ID NO: 51; andSEQ ID NO: 52) of the heavy chain variable region of SEQ ID NO: 38.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 53:

(SEQ ID NO: 61) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCATATGTGACCCTGTGCTGACCCAGACTCCATCTCCCGTATCTGCACCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGGCCAGTCAGAGTGTTTATGAGAACAACTATTTATCCTGGTTTCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGATTCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATTACAGACGTGCAGTGTGACGATGCTGCCACTTACTATTGTGCAGGCGTTTATGATGATGATAGTGATGATGCCTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 54:

(SEQ ID NO: 62) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTGGCTGTGCTCAAAGGTGTCCAGTGTCAGGAGCAGCTGAAGGAGTCCGGAGGAGGCCTGGTAACGCCTGGAGGAACCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAATGCCTACTACATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGATTCATTACTCTGAATAATAATGTAGCTTACGCGAACTGGGCGAAAGGCCGATTCACCTTCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACACCCGAGGACACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTGCAATGGGTCGGTTGGATCTCTGGGGCCATGGCACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAA GG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 63; SEQ ID NO: 64; and SEQ ID NO: 65 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 53.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 66; SEQ ID NO: 67; and SEQ ID NO: 68 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 54.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 61 encoding the light chainvariable region of SEQ ID NO: 53; the polynucleotide SEQ ID NO: 62encoding the heavy chain variable region of SEQ ID NO: 54;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 63; SEQ ID NO: 64; and SEQ ID NO: 65) of the light chain variableregion of SEQ ID NO: 53; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 66; SEQ ID NO: 67; andSEQ ID NO: 68) of the heavy chain variable region of SEQ ID NO: 54.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 69:

(SEQ ID NO: 77) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCCCAAGTGCTGACCCAGACTCCATCGCCTGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAACTGCCAGGCCAGTCAGAGTGTTGATGATAACAACTGGTTAGGCTGGTATCAGCAGAAACGAGGGCAGCCTCCCAAGTACCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCTGCCACTTACTACTGTGCAGGCGGTTTTAGTGGTAATATCTTTGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 70:

(SEQ ID NO: 78) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGCTTCTCCCTCAGTAGCTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGAAAGGGGCTGGAGTGGATCGGAATCATTGGTGGTTTTGGTACCACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAGAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGTGGTCCTGGTAATGGTGGTGACATCTGGGGCCAAGGGACCCTGGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACT.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 79; SEQ ID NO: 80; and SEQ ID NO: 81 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 69.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 82; SEQ ID NO: 83; and SEQ ID NO: 84 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 70.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 77 encoding the light chainvariable region of SEQ ID NO: 69; the polynucleotide SEQ ID NO: 78encoding the heavy chain variable region of SEQ ID NO: 70;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 79; SEQ ID NO: 80; and SEQ ID NO: 81) of the light chain variableregion of SEQ ID NO: 69; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 82; SEQ ID NO: 83; andSEQ ID NO: 84) of the heavy chain variable region of SEQ ID NO: 70.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 85:

(SEQ ID NO: 93) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGTACCTGTGGGAGGCACAGTCACCATCAAGTGCCAGTCCAGTCAGAGTGTTTATAATAATTTCTTATCGTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACCAGGCATCCAAACTGGCATCTGGGGTCCCAGATAGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTC

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 86:

(SEQ ID NO: 94) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACGCTCACCTGCACAGTCTCTGGAATCGACCTCAGTGACTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAATCATTTATGCTGGTAGTGGTAGCACATGGTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGATGGATACGATGACTATGGTGATTTCGATCGATTGGATCTCTGGGGCCCAGGCACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCT GGTCAAGGACT.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 95; SEQ ID NO: 96; and SEQ ID NO: 97 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the light chain variable sequence of SEQ IDNO: 85.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 98; SEQ ID NO: 99; and SEQ ID NO: 100 which correspond topolynucleotides encoding the complementarity-determining regions (CDRs,or hypervariable regions) of the heavy chain variable sequence of SEQ IDNO: 86.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 93 encoding the light chainvariable region of SEQ ID NO: 85; the polynucleotide SEQ ID NO: 94encoding the heavy chain variable region of SEQ ID NO: 86;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 95; SEQ ID NO: 96; and SEQ ID NO: 97) of the light chain variableregion of SEQ ID NO: 85; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 98; SEQ ID NO: 99; andSEQ ID NO: 100) of the heavy chain variable region of SEQ ID NO: 86.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 101:

(SEQ ID NO: 109) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCGGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAAATGCCAGGCCAGTCAGAGCATTAACAATGAATTATCCTGGTATCAGCAGAAATCAGGGCAGCGTCCCAAGCTCCTGATCTATAGGGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAACAGGGTTATAGTCTGAGGAATATTGATAATGCTTTCGGCGGAGGGACCGAGGTGGTGGTCAAACGTACGGTAGCGGCCCCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTC

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 102:

(SEQ ID NO: 110) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCTCAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTAACTACTACATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAATGATTTATGGTAGTGATGAAACAGCCTACGCGAACTGGGCGATAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCCAGAGATGATAGTAGTGACTGGGATGCAAAATTTAACTTGTGGGGCCAAGGGACCCTCGTCACCGTCTCGAGCGCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 111; SEQ ID NO: 112; and SEQ ID NO: 113 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 101.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 114; SEQ ID NO: 115; and SEQ ID NO: 116 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 102.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 109 encoding the light chainvariable region of SEQ ID NO: 101; the polynucleotide SEQ ID NO: 110encoding the heavy chain variable region of SEQ ID NO: 102;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 111; SEQ ID NO: 112; and SEQ ID NO: 113) of the light chain variableregion of SEQ ID NO: 101; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 114; SEQ ID NO: 115; andSEQ ID NO: 116) of the heavy chain variable region of SEQ ID NO: 102.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 122:

(SEQ ID NO: 130) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCAGCCGTGCTGACCCAGACACCATCACCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAGTTGCCAGTCCAGTCAGAGTGTTGGTAATAACCAGGACTTATCCTGGTTTCAGCAGAGACCAGGGCAGCCTCCCAAGCTCCTGATCTACGAAATATCCAAACTGGAATCTGGGGTCCCATCGCGGTTCAGCGGCAGTGGATCTGGGACACACTTCACTCTCACCATCAGCGGCGTACAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGGTTATGA TGATGATGCTGATAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 123:

(SEQ ID NO: 131) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCACTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAGTAGTCGTACAATGTCCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGATACATTTGGAGTGGTGGTAGCACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGATTGGGCGATACTGGTGGTCACGCTTATGCTACTCGCTTAAATCTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 132; SEQ ID NO: 133; and SEQ ID NO: 134 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 122.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 135; SEQ ID NO: 136; and SEQ ID NO: 137 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 123.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 130 encoding the light chainvariable region of SEQ ID NO: 122; the polynucleotide SEQ ID NO: 131encoding the heavy chain variable region of SEQ ID NO: 123;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 132; SEQ ID NO: 133; and SEQ ID NO: 134) of the light chain variableregion of SEQ ID NO: 122; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 135; SEQ ID NO: 136; andSEQ ID NO: 137) of the heavy chain variable region of SEQ ID NO: 123.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 138:

(SEQ ID NO: 146) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCAGCCGTGCTGACCCAGACACCATCGTCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGTCCAGTCAGAGTGTTTATAGTAATAAGTACCTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACTGGACATCCAAACTGGCATCTGGGGCCCCATCACGGTTCAGCGGCAGTGGATCTGGGACACAATTCACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGCTTATGA TGATGATGCTGATAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 139:

(SEQ ID NO: 147) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAAGAGTCCGGGGGTCGCCTGGTCAAGCCTGACGAAACCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTGGAGGGCGGCTACATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAATCAGTTATGATAGTGGTAGCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAGACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGCGTCAGATCACTAAAATATCCTACTGTTACTTCTGATGACTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 148; SEQ ID NO: 149; and SEQ ID NO: 150 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 138.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 151; SEQ ID NO: 152; and SEQ ID NO: 153 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 139.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 146 encoding the light chainvariable region of SEQ ID NO: 138; the polynucleotide SEQ ID NO: 147encoding the heavy chain variable region of SEQ ID NO: 139;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 148; SEQ ID NO: 149; and SEQ ID NO: 150) of the light chain variableregion of SEQ ID NO: 138; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 151; SEQ ID NO: 152; andSEQ ID NO: 153) of the heavy chain variable region of SEQ ID NO: 139.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 154:

(SEQ ID NO: 162) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCAGCCGTGCTGACCCAGACACCATCACCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAGTTGCCAGTCCAGTCAGAGTGTTTATAATAATAACGACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCTAAACTCCTGATCTATTATGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCGCTTACTACTGTCTAGGCGGTTATGATGATGATGCTGATAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 155:

(SEQ ID NO: 163) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTATCTGGATTATCCCTCAGTAGCAATACAATAAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGATACATTTGGAGTGGTGGTAGTACATACTACGCGAGCTGGGTGAATGGTCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGGTTACGCTAGTGGTGGTTATCCTTATGCCACTCGGTTGGATCTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 164; SEQ ID NO: 165; and SEQ ID NO: 166 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 154.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 167; SEQ ID NO: 168; and SEQ ID NO: 169 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 155.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 162 encoding the light chainvariable region of SEQ ID NO: 154; the polynucleotide SEQ ID NO: 163encoding the heavy chain variable region of SEQ ID NO: 155;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 164; SEQ ID NO: 165; and SEQ ID NO: 166) of the light chain variableregion of SEQ ID NO: 154; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 167; SEQ ID NO: 168; andSEQ ID NO: 169) of the heavy chain variable region of SEQ ID NO: 155.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 170:

(SEQ ID NO: 178) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCAGCCGTGCTGACCCAGACACCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGTCCAGTCAGAGTGTTTATAATAACGACTACTTATCCTGGTATCAACAGAGGCCAGGGCAACGTCCCAAGCTCCTAATCTATGGTGCTTCCAAACTGGCATCTGGGGTCCCGTCACGGTTCAAAGGCAGTGGATCTGGGAAACAGTTTACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTGGGCGATTATGATGATGATGCTGATAATACT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 171:

(SEQ ID NO: 179) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACTTGCACAGTCTCTGGATTCACCCTCAGTACCAACTACTACCTGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTAGAATGGATCGGAATCATTTATCCTAGTGGTAACACATATTGCGCGAAGTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGACACAGCCACGTATTTCTGTGCCAGAAATTATGGTGGTGATGAAAGTTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 180; SEQ ID NO: 181; and SEQ ID NO: 182 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 170.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 183; SEQ ID NO: 184; and SEQ ID NO: 185 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 171.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 178 encoding the light chainvariable region of SEQ ID NO: 170; the polynucleotide SEQ ID NO: 179encoding the heavy chain variable region of SEQ ID NO: 171;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 180; SEQ ID NO: 181; and SEQ ID NO: 182) of the light chain variableregion of SEQ ID NO: 170; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 183; SEQ ID NO: 184; andSEQ ID NO: 185) of the heavy chain variable region of SEQ ID NO: 171.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 186:

(SEQ ID NO: 194) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGACCATTGGCAATGCATTAGCCTGGTATCAGCAGAAATCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCAAACTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAATGGTGTTATTTTG GTGATAGTGTT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 187:

(SEQ ID NO: 195) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCACTGTGCTCAAAGGTGTCCAGTGTCAGGAGCAGCTGGTGGAGTCCGGGGGAGGCCTGGTCCAGCCTGAGGGATCCCTGACACTCACCTGCACAGCCTCTGGATTCGACTTCAGTAGCGGCTACTACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCGTGTATTTTCACTATTACTACTAACACTTACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAGACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTGACAGCCGCGGACACGGCCACCTATCTCTGTGCGAGAGGGATTTATTCTGATAATAATTATTATGCCTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 196; SEQ ID NO: 197; and SEQ ID NO: 198 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 186.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 199; SEQ ID NO: 200; and SEQ ID NO: 201 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 187.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 194 encoding the light chainvariable region of SEQ ID NO: 186; the polynucleotide SEQ ID NO: 195encoding the heavy chain variable region of SEQ ID NO: 187;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 196; SEQ ID NO: 197; and SEQ ID NO: 198) of the light chain variableregion of SEQ ID NO: 186; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 199; SEQ ID NO: 200; andSEQ ID NO: 201) of the heavy chain variable region of SEQ ID NO: 187.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 202:

(SEQ ID NO: 210) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGAGCATTGGCAATGCATTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGCCGATGCTGCCGCTTACTACTGTCAATGGTGTTATTTTG GTGATAGTGTT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 203:

(SEQ ID NO: 211) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGCAGCAGCTGGTGGAGTCCGGGGGAGGCCTGGTCAAGCCGGGGGCATCCCTGACACTCACCTGCAAAGCCTCTGGATTCTCCTTCAGTAGCGGCTACTACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTCGATCGCATGCATTTTTACTATTACTGATAACACTTACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAGCCCTCGTCGCCCACGGTGACTCTGCAAATGACCAGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAGGGGGATTTATTCTACTGATAATTATTATGCCTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 212; SEQ ID NO: 213; and SEQ ID NO: 214 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 202.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 215; SEQ ID NO: 216; and SEQ ID NO: 217 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 203.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 210 encoding the light chainvariable region of SEQ ID NO: 202; the polynucleotide SEQ ID NO: 211encoding the heavy chain variable region of SEQ ID NO: 203;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 212; SEQ ID NO: 213; and SEQ ID NO: 214) of the light chain variableregion of SEQ ID NO: 202; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 215; SEQ ID NO: 216; andSEQ ID NO: 217) of the heavy chain variable region of SEQ ID NO: 203.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 218:

(SEQ ID NO: 226) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGAGCGTTAGTAGCTACTTAAACTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAGGGCATCCACTCTGGAATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAATGTACTTATGGTACTAGTAGTAGTTATGGTGCTGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 219:

(SEQ ID NO: 227) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACCGTCTCTGGTATCTCCCTCAGTAGCAATGCAATAAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAATCATTAGTTATAGTGGTACCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGGATCTGAAAATCACTAGTCCGACAACCGAGGACACGGCCACCTACTTCTGTGCCAGAGATGACCCTACGACAGTTATGGTTATGTTGATACCTTTTGGAGCCGGCATGGAC CTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 228; SEQ ID NO: 229; and SEQ ID NO: 230 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 218.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 231; SEQ ID NO: 232; and SEQ ID NO: 233 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 219.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 226 encoding the light chainvariable region of SEQ ID NO: 218; the polynucleotide SEQ ID NO: 227encoding the heavy chain variable region of SEQ ID NO: 219;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 228; SEQ ID NO: 229; and SEQ ID NO: 230) of the light chain variableregion of SEQ ID NO: 218; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 231; SEQ ID NO: 232; andSEQ ID NO: 233) of the heavy chain variable region of SEQ ID NO: 219.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 234:

(SEQ ID NO: 242) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCCCAAGTGCTGACCCAGACTGCATCGCCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAACTGCCAGGCCAGTCAGAGTGTTTATAAGAACAACTACTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAAGGCCTGATCTATTCTGCATCGACTCTAGATTCTGGGGTCCCATTGCGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCAGTTATGATTGTAGTAGTGGTGATTGTTATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 235:

(SEQ ID NO: 243) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGTTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGAGGGATCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCTTCAGTAGCTACTGGATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGTTACTGGTAATGGTAACACTTACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTGACAGCCGCGGACACGGCCACCTATTTTTGTGCGAAAG CCTATGACTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 244; SEQ ID NO: 245; and SEQ ID NO: 246 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 234.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 247; SEQ ID NO: 248; and SEQ ID NO: 249 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 235.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 242 encoding the light chainvariable region of SEQ ID NO: 234; the polynucleotide SEQ ID NO: 243encoding the heavy chain variable region of SEQ ID NO: 235;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 244; SEQ ID NO: 245; and SEQ ID NO: 246) of the light chain variableregion of SEQ ID NO: 234; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 247; SEQ ID NO: 248; andSEQ ID NO: 249) of the heavy chain variable region of SEQ ID NO: 235.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 250:

(SEQ ID NO: 258) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTTCCACATTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGGCCAGTCAGAGTGTTTATGACAACAACTATTTATCCTGGTATCAGCAGAAACCAGGACAGCCTCCCAAGCTCCTGATCTATGGTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCACGGGATCTGGGACACAGTTCACTCTCACCATCACAGACGTGCAGTGTGACGATGCTGCCACTTACTATTGTGCAGGCGTTTTTAATGATGATAGTGATGATGCC

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 251:

(SEQ ID NO: 259) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCCCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACACTCTCTGGATTCTCCCTCAGTGCATACTATATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGATTCATTACTCTGAGTGATCATATATCTTACGCGAGGTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGGAGTCGTGGCTGGGGTGCAATGGGTCGGTTGGATCTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 260; SEQ ID NO: 261; and SEQ ID NO: 262 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 250.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 263; SEQ ID NO: 264; and SEQ ID NO: 265 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 251.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 258 encoding the light chainvariable region of SEQ ID NO: 250; the polynucleotide SEQ ID NO: 259encoding the heavy chain variable region of SEQ ID NO: 251;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 260; SEQ ID NO: 261; and SEQ ID NO: 262) of the light chain variableregion of SEQ ID NO: 250; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 263; SEQ ID NO: 264; andSEQ ID NO: 265) of the heavy chain variable region of SEQ ID NO: 251.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 266:

(SEQ ID NO: 274) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTCGCAGCCGTGCTGACCCAGACACCATCGCCCGTGTCTGCGGCTGTGGGAGGCACAGTCACCATCAGTTGCCAGGCCAGTCAGAGTGTTTATAACAACAAAAATTTAGCCTGGTATCAGCAGAAATCAGGGCAGCCTCCCAAGCTCCTGATCTACTGGGCATCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCGTCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGTTTTTGA TGATGATGCTGATAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 267:

(SEQ ID NO: 275) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAATGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTAGCTACTCCATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATATATCGGAGTCATTGGTACTAGTGGTAGCACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAGAACCTCGACCACGGTGGCTCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGTCAGGAGTCTTTCTTC TATTACTTTCTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 276; SEQ ID NO: 277; and SEQ ID NO: 278 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 266.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 279; SEQ ID NO: 280; and SEQ ID NO: 281 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 267.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 274 encoding the light chainvariable region of SEQ ID NO: 266; the polynucleotide SEQ ID NO: 275encoding the heavy chain variable region of SEQ ID NO: 267;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 276; SEQ ID NO: 277; and SEQ ID NO: 278) of the light chain variableregion of SEQ ID NO: 266; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 279; SEQ ID NO: 280; andSEQ ID NO: 281) of the heavy chain variable region of SEQ ID NO: 267.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 282:

(SEQ ID NO: 290) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCATTCGAATTGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTCAGAACATTTATAGATACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGTTCCTGATCTATCTGGCATCTACTCTGGCATCTGGGGTCCCATCGCGGTTTAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAAAGTTATTATAGTAGTAA TAGTGTCGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 283:

(SEQ ID NO: 291) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGGAGCAGCTGGTGGAGTCCGGGGGAGACCTGGTCCAGCCTGAGGGATCCCTGACACTCACCTGCACAGCTTCTGAGTTAGACTTCAGTAGCGGCTACTGGATATGCTGGGTCCGCCAGGTTCCAGGGAAGGGGCTGGAGTGGATCGGATGCATTTATACTGGTAGTAGTGGTAGCACTTTTTACGCGAGTTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAGAGGTTATAGTGGCTTTGGTTACTTTAAGTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 292; SEQ ID NO: 293; and SEQ ID NO: 294 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 282.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 295; SEQ ID NO: 296; and SEQ ID NO: 297 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 283.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 290 encoding the light chainvariable region of SEQ ID NO: 282; the polynucleotide SEQ ID NO: 291encoding the heavy chain variable region of SEQ ID NO: 283;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 292; SEQ ID NO: 293; and SEQ ID NO: 294) of the light chain variableregion of SEQ ID NO: 282; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 295; SEQ ID NO: 296; andSEQ ID NO: 297) of the heavy chain variable region of SEQ ID NO: 283.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 298:

(SEQ ID NO: 306) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGGACATTTATAGGTTATTGGCCTGGTATCAACAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGATTCATCCGATCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCGCCATCAGCGGTGTGCAGTGTGACGATGCTGCCACTTACTACTGTCAACAGGCTTGGAGTTATAG TGATATTGATAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 299:

(SEQ ID NO: 307) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCGGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAGTAGCTACTACATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAATCATTACTACTAGTGGTAATACATTTTACGCGAGCTGGGCGAAAGGCCGGCTCACCATCTCCAGAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAACTTCTGATAT TTTTTATTATCGTAACTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 308; SEQ ID NO: 309; and SEQ ID NO: 310 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 298.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 311; SEQ ID NO: 312; and SEQ ID NO: 313 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 299.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 306 encoding the light chainvariable region of SEQ ID NO: 298; the polynucleotide SEQ ID NO: 307encoding the heavy chain variable region of SEQ ID NO: 299;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 308; SEQ ID NO: 309; and SEQ ID NO: 310) of the light chain variableregion of SEQ ID NO: 298; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 311; SEQ ID NO: 312; andSEQ ID NO: 313) of the heavy chain variable region of SEQ ID NO: 299.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 314:

(SEQ ID NO: 322) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACGTTTGCAGCCGTGCTGACCCAGACTGCATCACCCGTGTCTGCCGCTGTGGGAGCCACAGTCACCATCAACTGCCAGTCCAGTCAGAGTGTTTATAATGACATGGACTTAGCCTGGTTTCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGCAGTGTGACGATGCTGCCACTTACTACTGTCTAGGCGCTTTTGA TGATGATGCTGATAATACT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 315:

(SEQ ID NO: 323) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCACTAGGCATGCAATAACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGATGCATTTGGAGTGGTGGTAGCACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTCAGAATCACCAGTCCGACAACCGAGGACACGGCCACCTACTTCTGTGCCAGAGTCATTGGCGATACTGCTGGTTATGCTTATTTTACGGGGCTTGACTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 324; SEQ ID NO: 325; and SEQ ID NO: 326 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 314.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 327; SEQ ID NO: 328; and SEQ ID NO: 329 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 315.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 322 encoding the light chainvariable region of SEQ ID NO: 314; the polynucleotide SEQ ID NO: 323encoding the heavy chain variable region of SEQ ID NO: 315;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 324; SEQ ID NO: 325; and SEQ ID NO: 326) of the light chain variableregion of SEQ ID NO: 314; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 327; SEQ ID NO: 328; andSEQ ID NO: 329) of the heavy chain variable region of SEQ ID NO: 315.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 330:

(SEQ ID NO: 338) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGAGTGTTTATAATTGGTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATACTGCATCCAGTCTGGCATCTGGGGTCCCATCGCGGTTCAGTGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGGCGTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAACAGGGTTATACTAGTGA TGTTGATAATGTT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 331:

(SEQ ID NO: 339) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGGCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGAATCGACCTCAGTAGCTATGCAATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATACATCGGAATCATTAGTAGTAGTGGTAGCACATACTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCACAAGCCTCGTCGACCACGGTGGATCTGAAAATTACCAGTCCGACAACCGAGGACTCGGCCACATATTTCTGTGCCAGAGGGGGTGCTGGTAGTGGTGGTGTTTGGCTGCTTGATGGTTTTGATCCC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 340; SEQ ID NO: 341; and SEQ ID NO: 342 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 330.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 343; SEQ ID NO: 344; and SEQ ID NO: 345 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 331.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 338 encoding the light chainvariable region of SEQ ID NO: 330; the polynucleotide SEQ ID NO: 339encoding the heavy chain variable region of SEQ ID NO: 331;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 340; SEQ ID NO: 341; and SEQ ID NO: 342) of the light chain variableregion of SEQ ID NO: 330; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 343; SEQ ID NO: 344; andSEQ ID NO: 345) of the heavy chain variable region of SEQ ID NO: 331.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 346:

(SEQ ID NO: 354) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAAATGTGCCGATGTTGTGATGACCCAGACTCCAGCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTGAGAACATTTATAATTGGTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATACTGTAGGCGATCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTATTGTCAACAGGGTTATAGTAG TAGTTATGTTGATAATGTT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 347:

(SEQ ID NO: 355) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGGAGCAGCTGAAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAATGACTATGCAGTGGGCTGGTTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGATACATTCGTAGTAGTGGTACCACAGCCTACGCGACCTGGGCGAAAGGCCGATTCACCATCTCCGCTACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGGGTGCTGGTAGTAGTGGTGTGTGGATCCTTGATGGTTTTGCTCCC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 356; SEQ ID NO: 357; and SEQ ID NO: 358 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 346.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 359; SEQ ID NO: 360; and SEQ ID NO: 361 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 347.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 354 encoding the light chainvariable region of SEQ ID NO: 346; the polynucleotide SEQ ID NO: 355encoding the heavy chain variable region of SEQ ID NO: 347;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 356; SEQ ID NO: 357; and SEQ ID NO: 358) of the light chain variableregion of SEQ ID NO: 346; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 359; SEQ ID NO: 360; andSEQ ID NO: 361) of the heavy chain variable region of SEQ ID NO: 347.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 362:

(SEQ ID NO: 370) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCTCAAGTGCTGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTCAGAGTGTTTATCAGAACAACTACTTATCCTGGTTTCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCGGCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCTGCCACTTACTACTGTGCAGGCGCTTATAG GGATGTGGATTCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 363:

(SEQ ID NO: 371) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGTTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGGGGCATCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCTTTACTAGTACCTACTACATCTACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGTATTGATGCTGGTAGTAGTGGTAGCACTTACTACGCGACCTGGGTGAATGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAAATGGGATTATGGTGGTAATGTTGGTTGGGGTTATGACTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 372; SEQ ID NO: 373; and SEQ ID NO: 374 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 362.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 375; SEQ ID NO: 376; and SEQ ID NO: 377 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 363.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 370 encoding the light chainvariable region of SEQ ID NO: 362; the polynucleotide SEQ ID NO: 371encoding the heavy chain variable region of SEQ ID NO: 363;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 372; SEQ ID NO: 373; and SEQ ID NO: 374) of the light chain variableregion of SEQ ID NO: 362; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 375; SEQ ID NO: 376; andSEQ ID NO: 377) of the heavy chain variable region of SEQ ID NO: 363.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 378:

(SEQ ID NO: 386) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCATTCGAATTGACCCAGACTCCATCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTCAGAGCATTAGTAGTTACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGTTCCTGATCTACAGGGCGTCCACTCTGGCATCTGGGGTCCCATCGCGATTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTACTGTCAAAGCTATTATGATAGTGT TTCAAATCCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 379:

(SEQ ID NO: 387) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGTTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGAGGGATCCCTGACACTCACCTGCAAAGCCTCTGGACTCGACCTCGGTACCTACTGGTTCATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCTTGTATTTATACTGGTAGTAGTGGTTCCACTTTCTACGCGAGCTGGGTGAATGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACTCTGCAAATGACCAGTCTGACAGCCGCGGACACGGCCACTTATTTTTGTGCGAGAGGTTATAGTGGTTATGGTTATTTTAAGTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 388; SEQ ID NO: 389; and SEQ ID NO: 390 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 378.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 391; SEQ ID NO: 392; and SEQ ID NO: 393 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 379.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 386 encoding the light chainvariable region of SEQ ID NO: 378; the polynucleotide SEQ ID NO: 387encoding the heavy chain variable region of SEQ ID NO: 379;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 388; SEQ ID NO: 389; and SEQ ID NO: 390) of the light chain variableregion of SEQ ID NO: 378; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 391; SEQ ID NO: 392; andSEQ ID NO: 393) of the heavy chain variable region of SEQ ID NO: 379.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 394:

(SEQ ID NO: 402) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGTCACATTTGCCATCGAAATGACCCAGAGTCCATTCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGGCCAGTCAGAGTGTTTATAAGAACAACCAATTATCCTGGTATCAGCAGAAATCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCGGCTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTGCAGGCGCTATTACTGGTAGTATTGATACGGATGGT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 395:

(SEQ ID NO: 403) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGTTGGAGGAGTCCGGGGGAGACCTGGTCAAGCCTGGGGCATCCCTGACACTCACCTGCACAACTTCTGGATTCTCCTTCAGTAGCAGCTACTTCATTTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTTATGGTGGTGATGGCAGCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGTCGACCACGGTGACGCTGCAAATGACCAGTCTGACAGCCGCGGACACGGCCACCTATTTCTGTGCGAGAGAATGGGCATATAGTCAAGGTTATTTTGGTGCTTTTGATCTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 404; SEQ ID NO: 405; and SEQ ID NO: 406 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 394.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 407; SEQ ID NO: 408; and SEQ ID NO: 409 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 395.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 402 encoding the light chainvariable region of SEQ ID NO: 394; the polynucleotide SEQ ID NO: 403encoding the heavy chain variable region of SEQ ID NO: 395;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 404; SEQ ID NO: 405; and SEQ ID NO: 406) of the light chain variableregion of SEQ ID NO: 394; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 407; SEQ ID NO: 408; andSEQ ID NO: 409) of the heavy chain variable region of SEQ ID NO: 395.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 410:

(SEQ ID NO: 418) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGGAGGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGGATATTAGTAGCTACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGCTGCATCCAATCTGGAATCTGGGGTCTCATCGCGATTCAAAGGCAGTGGATCTGGGACAGAGTACACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACCTATTACTGTCAATGTACTTATGGTACTATTTCTATTAGTGATGGTAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 411:

(SEQ ID NO: 419) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAATGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAGTAGCTACTTCATGACCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTGGAATACATCGGATTCATTAATCCTGGTGGTAGCGCTTACTACGCGAGCTGGGTGAAAGGCCGATTCACCATCTCCAAGTCCTCGACCACGGTAGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGGGTTCTGATTGTTTCTTATGGAGCCTTTACCATC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 420; SEQ ID NO: 421; and SEQ ID NO: 422 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 410.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 423; SEQ ID NO: 424; and SEQ ID NO: 425 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 411.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 418 encoding the light chainvariable region of SEQ ID NO: 410; the polynucleotide SEQ ID NO: 419encoding the heavy chain variable region of SEQ ID NO: 411;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 420; SEQ ID NO: 421; and SEQ ID NO: 422) of the light chain variableregion of SEQ ID NO: 410; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 423; SEQ ID NO: 424; andSEQ ID NO: 425) of the heavy chain variable region of SEQ ID NO: 411.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 426:

(SEQ ID NO: 434) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGATGTTGTGATGACCCAGACTCCAGCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCAGTGAGGACATTGAAAGCTATCTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGAATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCGACCTGGAGTGTGCCGATGCTGCCACTTACTATTGTCAATGCACTTATGGTATTATTAGTATTAGTGATGGTAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 427:

(SEQ ID NO: 435) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTGTCTGGATTCTCCCTCAGTAGCTACTTCATGACCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTGGAATACATCGGATTCATGAATACTGGTGATAACGCATACTACGCGAGCTGGGCGAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGGGTTCTTGTTGTTGCTTATGGAGCCTTTAACATC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 436; SEQ ID NO: 437; and SEQ ID NO: 438 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 426.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 439; SEQ ID NO: 440; and SEQ ID NO: 441 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 427.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 434 encoding the light chainvariable region of SEQ ID NO: 426; the polynucleotide SEQ ID NO: 435encoding the heavy chain variable region of SEQ ID NO: 427;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 436; SEQ ID NO: 437; and SEQ ID NO: 438) of the light chain variableregion of SEQ ID NO: 426; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 439; SEQ ID NO: 440; andSEQ ID NO: 441) of the heavy chain variable region of SEQ ID NO: 427.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 442:

(SEQ ID NO: 450) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGAACCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGTCCAGTAAGAGTGTTATGAATAACAACTACTTAGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATGGTGCATCCAATCTGGCATCTGGGGTCCCATCACGGTTCAGCGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGCTGCCACTTACTACTGTCAAGGCGGTTATACTGGTTATAGTGATCATGGGACT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 443:

(SEQ ID NO: 451) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCAAGCCTGACGAAACCCTGACACTCACCTGCACAGTCTCTGGAATCGACCTCAGTAGCTATCCAATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGATTCATTAATACTGGTGGTACCATAGTCTACGCGAGCTGGGCAAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGCAGTTATGTTTCATCTGGTTATGCCTACTATTTTAATGTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 452; SEQ ID NO: 453; and SEQ ID NO: 454 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 442.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 455; SEQ ID NO: 456; and SEQ ID NO: 457 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 443.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 450 encoding the light chainvariable region of SEQ ID NO: 442; the polynucleotide SEQ ID NO: 451encoding the heavy chain variable region of SEQ ID NO: 443;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 452; SEQ ID NO: 453; and SEQ ID NO: 454) of the light chain variableregion of SEQ ID NO: 442; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 455; SEQ ID NO: 456; andSEQ ID NO: 457) of the heavy chain variable region of SEQ ID NO: 443.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 458:

(SEQ ID NO: 466) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCCGCCGTGCTGACCCAGACTCCATCTCCCGTGTCTGCAGCTGTGGGAGGCACAGTCAGCATCAGTTGCCAGTCCAGTCAGAGTGTTTATAATAACAACTGGTTATCCTGGTTTCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAAGGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACGTGCAGTGTGACGATGTTGCCACTTACTACTGTGCGGGCGGTTATCT TGATAGTGTTATT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 459:

(SEQ ID NO: 467) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAGTACCTATTCAATAAACTGGGTCCGCCAGGCTCCAGGGAAGGGCCTGGAATGGATCGGAATCATTGCTAATAGTGGTACCACATTCTACGCGAACTGGGCGAAAGGCCGATTCACCGTCTCCAAAACCTCGACCACGGTGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGAGAGTGGAATGTACAATGAATATGGTAAATTTAACATC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 468; SEQ ID NO: 469; and SEQ ID NO: 470 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 458.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 471; SEQ ID NO: 472; and SEQ ID NO: 473 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 459.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 466 encoding the light chainvariable region of SEQ ID NO: 458; the polynucleotide SEQ ID NO: 467encoding the heavy chain variable region of SEQ ID NO: 459;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 468; SEQ ID NO: 469; and SEQ ID NO: 470) of the light chain variableregion of SEQ ID NO: 458; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 471; SEQ ID NO: 472; andSEQ ID NO: 473) of the heavy chain variable region of SEQ ID NO: 459.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 474:

(SEQ ID NO: 482) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTCTGATATGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTGAGAACATTTATAGCTTTTTGGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTTCAAGGCTTCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCTGCCACTTACTACTGTCAACAGGGTGCTACTGTGTA TGATATTGATAATAAT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 475:

(SEQ ID NO: 483) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTTTCTGGAATCGACCTCAGTGCCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTGGAATGGATCACAATCATTTATCCTAATGGTATCACATACTACGCGAACTGGGCGAAAGGCCGATTCACCGTCTCCAAAACCTCGACCGCGATGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGATGCAGAAAGTAGTAAGAATGCTTATTGGGGCTACTTTAACGTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 484; SEQ ID NO: 485; and SEQ ID NO: 486 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 474.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 487; SEQ ID NO: 488; and SEQ ID NO: 489 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 475.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 482 encoding the light chainvariable region of SEQ ID NO: 474; the polynucleotide SEQ ID NO: 483encoding the heavy chain variable region of SEQ ID NO: 475;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 484; SEQ ID NO: 485; and SEQ ID NO: 486) of the light chain variableregion of SEQ ID NO: 474; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 487; SEQ ID NO: 488; andSEQ ID NO: 489) of the heavy chain variable region of SEQ ID NO: 475.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 490:

(SEQ ID NO: 498) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTCTGATATGACCCAGACTCCATCCTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTGAGAACATTTATAGCTTTTTGGCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTTCAGGGCTTCCACTCTGGCATCTGGGGTCTCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGACCTGGAGTGTGACGATGCTGCCACTTACTACTGTCAACAGGGTGCTACTGTGTA TGATATTGATAATAAT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 491:

(SEQ ID NO: 499) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTTTCTGGAATCGACCTCAGTGCCTATGCAATGATCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTGGAATGGATCACAATCATTTATCCTAATGGTATCACATACTACGCGAACTGGGCGAAAGGCCGATTCACCGTCTCCAAAACCTCGACCGCGATGGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGATGCAGAAAGTAGTAAGAATGCTTATTGGGGCTACTTTAACGTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 500; SEQ ID NO: 501; and SEQ ID NO: 502 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 490.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 503; SEQ ID NO: 504; and SEQ ID NO: 505 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 491.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 498 encoding the light chainvariable region of SEQ ID NO: 490; the polynucleotide SEQ ID NO: 499encoding the heavy chain variable region of SEQ ID NO: 491;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 500; SEQ ID NO: 501; and SEQ ID NO: 502) of the light chain variableregion of SEQ ID NO: 490; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 503; SEQ ID NO: 504; andSEQ ID NO: 505) of the heavy chain variable region of SEQ ID NO: 491.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 506:

(SEQ ID NO: 514) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCCATTGAAATGACCCAGACTCCATCCCCCGTGTCTGCCGCTGTGGGAGGCACAGTCACCATCAATTGCCAGGCCAGTGAGAGTGTTTTTAATAATATGTTATCCTGGTATCAGCAGAAACCAGGGCACTCTCCTAAGCTCCTGATCTATGATGCATCCGATCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGTGGCGTGGAGTGTGACGATGCTGCCACTTACTATTGTGCAGGGTATAAAAGTGATAGTAATGATGGCGATAATGTT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 507:

(SEQ ID NO: 515) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAACAGGAATTCAATAACCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTGGAATGGATCGGAATCATTACTGGTAGTGGTAGAACGTACTACGCGAACTGGGCAAAAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAAATGACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGCCATCCTGG TCTTGGTAGTGGTAACATC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 516; SEQ ID NO: 517; and SEQ ID NO: 518 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 506.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 519; SEQ ID NO: 520; and SEQ ID NO: 521 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 507.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 514 encoding the light chainvariable region of SEQ ID NO: 506; the polynucleotide SEQ ID NO: 515encoding the heavy chain variable region of SEQ ID NO: 507;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 516; SEQ ID NO: 517; and SEQ ID NO: 518) of the light chain variableregion of SEQ ID NO: 506; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 519; SEQ ID NO: 520; andSEQ ID NO: 521) of the heavy chain variable region of SEQ ID NO: 507.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 522:

(SEQ ID NO: 530) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCGCAAGTGCTGACCCAGACTGCATCGTCCGTGTCTGCAGCTGTGGGAGGCACAGTCACCATCAATTGCCAGTCCAGTCAGAGTGTTTATAATAACTACTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTATACTGCATCCAGCCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGAAGTGCAGTGTGACGATGCTGCCACTTACTACTGTCAAGGCTATTATAGTGG TCCTATAATTACT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 523:

(SEQ ID NO: 531) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGCCTCTGGATTCTCCCTCAATAACTACTACATACAATGGGTCCGCCAGGCTCCAGGGGAGGGGCTGGAATGGATCGGGATCATTTATGCTGGTGGTAGCGCATACTACGCGACCTGGGCAAACGGCCGATTCACCATCGCCAAAACCTCGTCGACCACGGTGGATCTGAAGATGACCAGTCTGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGAGGGACATT TGATGGTTATGAGTTG.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 532; SEQ ID NO: 533; and SEQ ID NO: 534 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 522.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 535; SEQ ID NO: 536; and SEQ ID NO: 537 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 523.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 530 encoding the light chainvariable region of SEQ ID NO: 522; the polynucleotide SEQ ID NO: 531encoding the heavy chain variable region of SEQ ID NO: 523;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 532; SEQ ID NO: 533; and SEQ ID NO: 534) of the light chain variableregion of SEQ ID NO: 522; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 535; SEQ ID NO: 536; andSEQ ID NO: 537) of the heavy chain variable region of SEQ ID NO: 523.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 538:

(SEQ ID NO: 546) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCACATTTGCCCAAGTGCTGACCCAGACTCCATCCCCTGTGTCTGTCCCTGTGGGAGACACAGTCACCATCAGTTGCCAGTCCAGTGAGAGCGTTTATAGTAATAACCTCTTATCCTGGTATCAGCAGAAACCAGGGCAGCCTCCCAAGCTCCTGATCTACAGGGCATCCAATCTGGCATCTGGTGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCAGCGGCGCACAGTGTGACGATGCTGCCACTTACTACTGTCAAGGCTATTATAG TGGTGTCATTAATAGT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 539:

(SEQ ID NO: 547) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGGTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACAGTGTCTGGATTCTCCCTCAGTAGCTACTTCATGAGCTGGGTCCGCCAGGCTCCAGGGGAGGGGCTGGAATACATCGGATTCATTAATCCTGGTGGTAGCGCATACTACGCGAGCTGGGCGAGTGGCCGACTCACCATCTCCAAAACCTCGACCACGGTAGATCTGAAAATCACCAGTCCGACAACCGAGGACACGGCCACCTATTTCTGTGCCAGGATTCTTATTGTTTCTTATGGAGCCTTTACCATC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 548; SEQ ID NO: 549; and SEQ ID NO: 550 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 538.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 551; SEQ ID NO: 552; and SEQ ID NO: 553 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 539.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 546 encoding the light chainvariable region of SEQ ID NO: 538; the polynucleotide SEQ ID NO: 547encoding the heavy chain variable region of SEQ ID NO: 539;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 548; SEQ ID NO: 549; and SEQ ID NO: 550) of the light chain variableregion of SEQ ID NO: 538; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 551; SEQ ID NO: 552; andSEQ ID NO: 553) of the heavy chain variable region of SEQ ID NO: 539.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 554:

(SEQ ID NO: 562) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCACTGAGAGCATTGGCAATGAGTTATCCTGGTATCAGCAGAAACCAGGGCAGGCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACCGGCGTGGAGTGTGATGATGCTGCCACTTACTACTGTCAACAGGGTTATAGTAGTGC TAATATTGATAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 555:

(SEQ ID NO: 563) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTCACGCCTGGGACACCCCTGACACTCACCTGCACCGTCTCTGGATTCTCCCTCAGTAAGTACTACATGAGCTGGGTCCGCCAGGCTCCAGAGAAGGGGCTGAAATACATCGGATACATTGATAGTACTACTGTTAATACATACTACGCGACCTGGGCGAGAGGCCGATTCACCATCTCCAAAACCTCGACCACGGTGGATCTGAAGATCACCAGTCCGACAAGTGAGGACACGGCCACCTATTTCTGTGCCAGAGGAAGTACTTATTTTACTGATGGAGGCCATCGGTTGGATCTC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 564; SEQ ID NO: 565; and SEQ ID NO: 566 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 554.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 567; SEQ ID NO: 568; and SEQ ID NO: 569 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 555.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 562 encoding the light chainvariable region of SEQ ID NO: 554; the polynucleotide SEQ ID NO: 563encoding the heavy chain variable region of SEQ ID NO: 555;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 564; SEQ ID NO: 565; and SEQ ID NO: 566) of the light chain variableregion of SEQ ID NO: 554; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 567; SEQ ID NO: 568; andSEQ ID NO: 569) of the heavy chain variable region of SEQ ID NO: 555.

The invention is further directed to polynucleotides encodingpolypeptides of the antibodies having binding specificity to IL-6. Inone embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable light chain polypeptide sequence of SEQID NO: 570:

(SEQ ID NO: 578) ATGGACACGAGGGCCCCCACTCAGCTGCTGGGGCTCCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCTATGATATGACCCAGACTCCAGCCTCTGTGGAGGTAGCTGTGGGAGGCACAGTCACCATCAAGTGCCAGGCCACTGAGAGCATTGGCAATGAGTTATCCTGGTATCAGCAGAAACCAGGGCAGGCTCCCAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCGCGGTTCAAAGGCAGTGGATCTGGGACACAGTTCACTCTCACCATCACCGGCGTGGAGTGTGATGATGCTGCCACTTACTACTGTCAACAGGGTTATAGTAGTGC TAATATTGATAATGCT

In another embodiment of the invention, polynucleotides of the inventioncomprise, or alternatively consist of, the following polynucleotidesequence encoding the variable heavy chain polypeptide sequence of SEQID NO: 571:

(SEQ ID NO: 579) ATGGAGACTGGGCTGCGCTGGCTTCTCCTGGTCGCTGTGCTCAAAGGTGTCCAGTGTCAGTCGCTGGAGGAGTCCGGGGGTCGCCTGGTAACGCCTGGGACACCCCTGACACTCACCTGCACAGTCTCTGGATTCTCCCTCAGTACCTACAACATGGGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAATGGATCGGAAGTATTACTATTGATGGTCGCACATACTACGCGAGCTGGGCGAAAGGCCGATTCACCGTCTCCAAAAGCTCGACCACGGTGGATCTGAAAATGACCAGTCTGACAACCGGGGACACGGCCACCTATTTCTGTGCCAGGATTCTTATTGTTTCTTATGGGGCCTTTACCATC.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 580; SEQ ID NO: 581; and SEQ ID NO: 582 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the light chain variable sequence ofSEQ ID NO: 570.

In a further embodiment of the invention, polynucleotides encodingfragments of the antibody having binding specificity to IL-6 comprise,or alternatively consist of, one or more of the polynucleotide sequencesof SEQ ID NO: 583; SEQ ID NO: 584; and SEQ ID NO: 585 which correspondto polynucleotides encoding the complementarity-determining regions(CDRs, or hypervariable regions) of the heavy chain variable sequence ofSEQ ID NO: 571.

The invention also contemplates polynucleotide sequences including oneor more of the polynucleotide sequences encoding antibody fragmentsdescribed herein. In one embodiment of the invention, polynucleotidesencoding fragments of the antibody having binding specificity to IL-6comprise, or alternatively consist of, one, two, three or more,including all of the following polynucleotides encoding antibodyfragments: the polynucleotide SEQ ID NO: 578 encoding the light chainvariable region of SEQ ID NO: 570; the polynucleotide SEQ ID NO: 579encoding the heavy chain variable region of SEQ ID NO: 571;polynucleotides encoding the complementarity-determining regions (SEQ IDNO: 580; SEQ ID NO: 581; and SEQ ID NO: 582) of the light chain variableregion of SEQ ID NO: 570; and polynucleotides encoding thecomplementarity-determining regions (SEQ ID NO: 583; SEQ ID NO: 584; andSEQ ID NO: 585) of the heavy chain variable region of SEQ ID NO: 571.

In another embodiment of the invention, polynucleotides of the inventionfurther comprise, the following polynucleotide sequence encoding thekappa constant light chain sequence of SEQ ID NO: 586:

(SEQ ID NO: 587) GTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTC AACAGGGGAGAGTGT.

In another embodiment of the invention, polynucleotides of the inventionfurther comprise, the following polynucleotide sequence encoding thegamma-1 constant heavy chain polypeptide sequence of SEQ ID NO: 588:

(SEQ ID NO: 589) GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACGCCAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA.

In one embodiment, the invention is directed to an isolatedpolynucleotide comprising a polynucleotide encoding an anti-IL-6 V_(H)antibody amino acid sequence selected from SEQ ID NO: 3, 18, 19, 22, 38,54, 70, 86, 102, 117, 118, 123, 139, 155, 171, 187, 203, 219, 235, 251,267, 283, 299, 315, 331, 347, 363, 379, 395, 411, 427, 443, 459, 475,491, 507, 523, 539, 555 and SEQ ID NO: 571 or encoding a variant thereofwherein at least one framework residue (FR residue) has been substitutedwith an amino acid present at the corresponding position in a rabbitanti-IL-6 antibody V_(H) polypeptide or a conservative amino acidsubstitution.

In another embodiment, the invention is directed to an isolatedpolynucleotide comprising the polynucleotide sequence encoding ananti-IL-6 V_(L) antibody amino acid sequence of 2, 20, 21, 37, 53, 69,85, 101, 119, 122, 138, 154, 170, 186, 202, 218, 234, 250, 266, 282,298, 314, 330, 346, 362, 378, 394, 410, 426, 442, 458, 474, 490, 506,522, 538, 554 and SEQ ID NO: 570 or encoding a variant thereof whereinat least one framework residue (FR residue) has been substituted with anamino acid present at the corresponding position in a rabbit anti-IL-6antibody V_(L) polypeptide or a conservative amino acid substitution.

In yet another embodiment, the invention is directed to one or moreheterologous polynucleotides comprising a sequence encoding thepolypeptides contained in SEQ ID NO:2 and SEQ ID NO:3; SEQ ID NO:2 andSEQ ID NO:18; SEQ ID NO:2 and SEQ ID NO:19; SEQ ID NO:20 and SEQ IDNO:3; SEQ ID NO:20 and SEQ ID NO:18; SEQ ID NO:20 and SEQ ID NO:19; SEQID NO:21 and SEQ ID NO:22; SEQ ID NO:37 and SEQ ID NO:38; SEQ ID NO:53and SEQ ID NO:54; SEQ ID NO:69 and SEQ ID NO:70; SEQ ID NO:85 and SEQ IDNO:86; SEQ ID NO:101 and SEQ ID NO:102; SEQ ID NO:101 and SEQ ID NO:117;SEQ ID NO:101 and SEQ ID NO:118; SEQ ID NO:119 and SEQ ID NO:102; SEQ IDNO:119 and SEQ ID NO:117; SEQ ID NO:119 and SEQ ID NO:118; SEQ ID NO:122and SEQ ID NO:123; SEQ ID NO:138 and SEQ ID NO:139; SEQ ID NO:154 andSEQ ID NO:155; SEQ ID NO:170 and SEQ ID NO:171; SEQ ID NO:186 and SEQ IDNO:187; SEQ ID NO:202 and SEQ ID NO:203; SEQ ID NO:218 and SEQ IDNO:219; SEQ ID NO:234 and SEQ ID NO:235; SEQ ID NO:250 and SEQ IDNO:251; SEQ ID NO:266 and SEQ ID NO:267; SEQ ID NO:282 and SEQ IDNO:283; SEQ ID NO:298 and SEQ ID NO:299; SEQ ID NO:314 and SEQ IDNO:315; SEQ ID NO:330 and SEQ ID NO:331; SEQ ID NO:346 and SEQ IDNO:347; SEQ ID NO:362 and SEQ ID NO:363; SEQ ID NO:378 and SEQ IDNO:379; SEQ ID NO:394 and SEQ ID NO:395; SEQ ID NO:410 and SEQ IDNO:411; SEQ ID NO:426 and SEQ ID NO:427; SEQ ID NO:442 and SEQ IDNO:443; SEQ ID NO:458 and SEQ ID NO:459; SEQ ID NO:474 and SEQ IDNO:475; SEQ ID NO:490 and SEQ ID NO:491; SEQ ID NO:506 and SEQ IDNO:507; SEQ ID NO:522 and SEQ ID NO:523; SEQ ID NO:538 and SEQ IDNO:539; SEQ ID NO:554 and SEQ ID NO:555; or SEQ ID NO:570 and SEQ IDNO:571.

In another embodiment, the invention is directed to an isolated isolatedpolynucleotide that expresses a polypeptide containing at least one CDRpolypeptide derived from an anti-IL-6 antibody wherein said expressedpolypeptide alone specifically binds IL-6 or specifically binds IL-6when expressed in association with another polynucleotide sequence thatexpresses a polypeptide containing at least one CDR polypeptide derivedfrom an anti-IL-6 antibody wherein said at least one CDR is selectedfrom those contained in the V_(L) or V_(H) polypeptides contained in SEQID NO: 3, 18, 19, 22, 38, 54, 70, 86, 102, 117, 118, 123, 139, 155, 171,187, 203, 219, 235, 251, 267, 283, 299, 315, 331, 347, 363, 379, 395,411, 427, 443, 459, 475, 491, 507, 523, 539, 555; 571; 2, 20, 21, 37,53, 69, 85, 101, 119, 122, 138, 154, 170, 186, 202, 218, 234, 250, 266,282, 298, 314, 330, 346, 362, 378, 394, 410, 426, 442, 458, 474, 490,506, 522, 538, 554 and SEQ ID NO: 570.

Host cells and vectors comprising said polynucleotides are alsocontemplated.

The invention further contemplates vectors comprising the polynucleotidesequences encoding the variable heavy and light chain polypeptidesequences, as well as the individual complementarity determining regions(CDRs, or hypervariable regions) set forth herein, as well as host cellscomprising said sequences. In one embodiment of the invention, the hostcell is a yeast cell. In another embodiment of the invention, the yeasthost cell belongs to the genus Pichia.

Anti-IL-6 Activity

As stated previously, IL-6 is a member of a family of cytokines thatpromote cellular responses through a receptor complex consisting of atleast one subunit of the signal-transducing glycoprotein gp130 and theIL-6 receptor (IL-6R). The IL-6R may also be present in a soluble form(sIL-6R). IL-6 binds to IL-6R, which then dimerizes thesignal-transducing receptor gp130.

It is believed that the anti-IL-6 antibodies of the invention, or IL-6binding fragments thereof, are useful by exhibiting anti-IL-6 activity.In one non-limiting embodiment of the invention, the anti-IL-6antibodies of the invention, or IL-6 binding fragments thereof, exhibitanti-IL-6 activity by binding to IL-6 which may be soluble IL-6 or cellsurface expressed IL-6 and/or may prevent or inhibit the binding of IL-6to IL-6R and/or activation (dimerization) of the gp130signal-transducing glycoprotein and the formation of IL-6/IL-6R/gp130multimers and the biological effects of any of the foregoing. Thesubject anti-IL-6 antibodies may possess different antagonisticactivities based on where (i.e., epitope) the particular antibody bindsIL-6 and/or how it affects the formation of the foregoing IL-6 complexesand/or multimers and the biological effects thereof. Consequently,different anti-IL-6 antibodies according to the invention e.g., may bebetter suited for preventing or treating conditions involving theformation and accumulation of substantial soluble IL-6 such asrheumatoid arthritis whereas other antibodies may be favored intreatments wherein the prevention of IL-6/IL-6R/gp130 orIL-6/IL-6R/gp130 multimers is a desired therapeutic outcome. This can bedetermined in binding and other assays.

The anti-IL-6 activity of the anti-IL-6 antibody of the presentinvention, and fragments thereof having binding specificity to IL-6, mayalso be described by their strength of binding or their affinity forIL-6. This also may affect their therapeutic properties. In oneembodiment of the invention, the anti-IL-6 antibodies of the presentinvention, and fragments thereof having binding specificity to IL-6,bind to IL-6 with a dissociation constant (K_(D)) of less than or equalto 5×10⁻⁷, 10⁻⁷, 5×10⁻⁸, 10⁻⁸, 5×10⁻⁹, 10⁻⁹, 5×10⁻¹⁰, 10⁻¹⁰, 5×10⁻¹¹,10⁻¹¹, 5×10⁻¹², 10⁻¹², 5×10⁻¹³, 10⁻¹³, 5×10⁻¹⁴, 10⁻¹⁴, 5×10⁻¹⁵ or 10⁻¹⁵,Preferably, the anti-IL-6 antibodies and fragments thereof bind IL-6with a dissociation constant of less than or equal to 5×10⁻¹⁰.

In another embodiment of the invention, the anti-IL-6 activity of theanti-IL-6 antibodies of the present invention, and fragments thereofhaving binding specificity to IL-6, bind to IL-6 with an off-rate ofless than or equal to 10⁻⁴ S⁻¹, 5×10⁻⁵ S⁻¹, 10⁻⁵ S⁻¹, 5×10⁻⁶ S⁻¹, 10⁻⁶S⁻¹, 5×10⁻⁷ S⁻¹, or 10⁻⁷ S⁻¹. In one embodiment of the invention, theanti-IL-6 antibodies of the invention, and fragments thereof havingbinding specificity to IL-6, bind to a linear or conformational IL-6epitope.

In a further embodiment of the invention, the anti-IL-6 activity of theanti-IL-6 antibodies of the present invention, and fragments thereofhaving binding specificity to IL-6, exhibit anti-IL-6 activity byameliorating or reducing the symptoms of, or alternatively treating, orpreventing, diseases and disorders associated with IL-6. Non-limitingexamples of diseases and disorders associated with IL-6 are set forthinfra. As noted cancer-related fatigue, cachexia and rheumatoidarthritis are preferred indications for the subject anti-IL-6antibodies.

In another embodiment of the invention, the anti-IL-6 antibodiesdescribed herein, or IL-6 binding fragments thereof, do not have bindingspecificity for IL-6R or the gp-130 signal-transducing glycoprotein.

B-Cell Screening and Isolation

In one embodiment, the present invention provides methods of isolating aclonal population of antigen-specific B cells that may be used forisolating at least one antigen-specific cell. As described andexemplified infra, these methods contain a series of culture andselection steps that can be used separately, in combination,sequentially, repetitively, or periodically. Preferably, these methodsare used for isolating at least one antigen-specific cell, which can beused to produce a monoclonal antibody, which is specific to a desiredantigen, or a nucleic acid sequence corresponding to such an antibody.

In one embodiment, the present invention provides a method comprisingthe steps of:

a. preparing a cell population comprising at least one antigen-specificB cell;

b. enriching the cell population, e.g., by chromatography, to form anenriched cell population comprising at least one antigen-specific Bcell;

c. isolating a single B cell from the enriched B cell population; and

d. determining whether the single B cell produces an antibody specificto the antigen.

In another embodiment, the present invention provides an improvement toa method of isolating a single, antibody-producing B cell, theimprovement comprising enriching a B cell population obtained from ahost that has been immunized or naturally exposed to an antigen, whereinthe enriching step precedes any selection steps, comprises at least oneculturing step, and results in a clonal population of B cells thatproduces a single monoclonal antibody specific to said antigen.

Throughout this application, a “clonal population of B cells” refers toa population of B cells that only secrete a single antibody specific toa desired antigen. That is to say that these cells produce only one typeof monoclonal antibody specific to the desired antigen.

In the present application, “enriching” a cell population cells meansincreasing the frequency of desired cells, typically antigen-specificcells, contained in a mixed cell population, e.g., a B cell-containingisolate derived from a host that is immunized against a desired antigen.Thus, an enriched cell population encompasses a cell population having ahigher frequency of antigen-specific cells as a result of an enrichmentstep, but this population of cells may contain and produce differentantibodies.

The general term “cell population” encompasses pre- and apost-enrichment cell populations, keeping in mind that when multipleenrichment steps are performed, a cell population can be both pre- andpost-enrichment. For example, in one embodiment, the present inventionprovides a method:

a. harvesting a cell population from an immunized host to obtain aharvested cell population;

b. creating at least one single cell suspension from the harvested cellpopulation;

c. enriching at least one single cell suspension to form a firstenriched cell population;

d. enriching the first enriched cell population to form a secondenriched cell population;

e. enriching the second enriched cell population to form a thirdenriched cell population; and

f. selecting an antibody produced by an antigen-specific cell of thethird enriched cell population.

Each cell population may be used directly in the next step, or it can bepartially or wholly frozen for long- or short-term storage or for latersteps. Also, cells from a cell population can be individually suspendedto yield single cell suspensions. The single cell suspension can beenriched, such that a single cell suspension serves as thepre-enrichment cell population. Then, one or more antigen-specificsingle cell suspensions together form the enriched cell population; theantigen-specific single cell suspensions can be grouped together, e.g.,re-plated for further analysis and/or antibody production.

In one embodiment, the present invention provides a method of enrichinga cell population to yield an enriched cell population having anantigen-specific cell frequency that is about 50% to about 100%, orincrements therein. Preferably, the enriched cell population has anantigen-specific cell frequency greater than or equal to about 50%, 60%,70%, 75%, 80%, 90%, 95%, 99%, or 100%.

In another embodiment, the present invention provides a method ofenriching a cell population whereby the frequency of antigen-specificcells is increased by at least about 2-fold, 5-fold, 10-fold, 20-fold,50-fold, 100-fold, or increments therein.

Throughout this application, the term “increment” is used to define anumerical value in varying degrees of precision, e.g., to the nearest10, 1, 0.1, 0.01, etc. The increment can be rounded to any measurabledegree of precision, and the increment need not be rounded to the samedegree of precision on both sides of a range. For example, the range 1to 100 or increments therein includes ranges such as 20 to 80, 5 to 50,and 0.4 to 98. When a range is open-ended, e.g., a range of less than100, increments therein means increments between 100 and the measurablelimit. For example, less than 100 or increments therein means 0 to 100or increments therein unless the feature, e.g., temperature, is notlimited by 0.

Antigen-specificity can be measured with respect to any antigen. Theantigen can be any substance to which an antibody can bind including,but not limited to, peptides, proteins or fragments thereof,carbohydrates; organic and inorganic molecules; receptors produced byanimal cells, bacterial cells, and viruses; enzymes; agonists andantagonists of biological pathways; hormones; and cytokines. Exemplaryantigens include, but are not limited to, IL-2, IL-4, IL-6, IL-10,IL-12, IL-13, IL-18, IFN-α, IFN-γ, BAFF, CXCL13, IP-10, VEGF, EPO, EGF,HRG, Hepatocyte Growth Factor (HGF) and Hepcidin. Preferred antigensinclude IL-6, IL-13, TNF-α, VEGF-α, Hepatocyte Growth Factor (HGF) andHepcidin. In a method utilizing more than one enrichment step, theantigen used in each enrichment step can be the same as or differentfrom one another. Multiple enrichment steps with the same antigen mayyield a large and/or diverse population of antigen-specific cells;multiple enrichment steps with different antigens may yield an enrichedcell population with cross-specificity to the different antigens.

Enriching a cell population can be performed by any cell-selection meansknown in the art for isolating antigen-specific cells. For example, acell population can be enriched by chromatographic techniques, e.g.,Miltenyi bead or magnetic bead technology. The beads can be directly orindirectly attached to the antigen of interest. In a preferredembodiment, the method of enriching a cell population includes at leastone chromatographic enrichment step.

A cell population can also be enriched by performed by anyantigen-specificity assay technique known in the art, e.g., an ELISAassay or a halo assay. ELISA assays include, but are not limited to,selective antigen immobilization (e.g., biotinylated antigen capture bystreptavidin, avidin, or neutravidin coated plate), non-specific antigenplate coating, and through an antigen build-up strategy (e.g., selectiveantigen capture followed by binding partner addition to generate aheteromeric protein-antigen complex). The antigen can be directly orindirectly attached to a solid matrix or support, e.g., a column. A haloassay comprises contacting the cells with antigen-loaded beads andlabeled anti-host antibody specific to the host used to harvest the Bcells. The label can be, e.g., a fluorophore. In one embodiment, atleast one assay enrichment step is performed on at least one single cellsuspension. In another embodiment, the method of enriching a cellpopulation includes at least one chromatographic enrichment step and atleast one assay enrichment step.

Methods of “enriching” a cell population by size or density are known inthe art. See, e.g., U.S. Pat. No. 5,627,052. These steps can be used inthe present method in addition to enriching the cell population byantigen-specificity.

The cell populations of the present invention contain at least one cellcapable of recognizing an antigen. Antigen-recognizing cells include,but are not limited to, B cells, plasma cells, and progeny thereof. Inone embodiment, the present invention provides a clonal cell populationcontaining a single type of antigen-specific B-cell, i.e., the cellpopulation produces a single monoclonal antibody specific to a desiredantigen.

In such embodiment, it is believed that the clonal antigen-specificpopulation of B cells consists predominantly of antigen-specific,antibody-secreting cells, which are obtained by the novel culture andselection protocol provided herein. Accordingly, the present inventionalso provides methods for obtaining an enriched cell populationcontaining at least one antigen-specific, antibody-secreting cell. Inone embodiment, the present invention provides an enriched cellpopulation containing about 50% to about 100%, or increments therein, orgreater than or equal to about 60%, 70%, 80%, 90%, or 100% ofantigen-specific, antibody-secreting cells.

In one embodiment, the present invention provides a method of isolatinga single B cell by enriching a cell population obtained from a hostbefore any selection steps, e.g., selecting a particular B cell from acell population and/or selecting an antibody produced by a particularcell. The enrichment step can be performed as one, two, three, or moresteps. In one embodiment, a single B cell is isolated from an enrichedcell population before confirming whether the single B cell secretes anantibody with antigen-specificity and/or a desired property.

In one embodiment, a method of enriching a cell population is used in amethod for antibody production and/or selection. Thus, the presentinvention provides a method comprising enriching a cell populationbefore selecting an antibody. The method can include the steps of:preparing a cell population comprising at least one antigen-specificcell, enriching the cell population by isolating at least oneantigen-specific cell to form an enriched cell population, and inducingantibody production from at least one antigen-specific cell. In apreferred embodiment, the enriched cell population contains more thanone antigen-specific cell. In one embodiment, each antigen-specific cellof the enriched population is cultured under conditions that yield aclonal antigen-specific B cell population before isolating an antibodyproducing cell therefrom and/or producing an antibody using said B cell,or a nucleic acid sequence corresponding to such an antibody. Incontrast to prior techniques where antibodies are produced from a cellpopulation with a low frequency of antigen-specific cells, the presentinvention allows antibody selection from among a high frequency ofantigen-specific cells. Because an enrichment step is used prior toantibody selection, the majority of the cells, preferably virtually allof the cells, used for antibody production are antigen-specific. Byproducing antibodies from a population of cells with an increasedfrequency of antigen specificity, the quantity and variety of antibodiesare increased.

In the antibody selection methods of the present invention, an antibodyis preferably selected after an enrichment step and a culture step thatresults in a clonal population of antigen-specific B cells. The methodscan further comprise a step of sequencing a selected antibody orportions thereof from one or more isolated, antigen-specific cells. Anymethod known in the art for sequencing can be employed and can includesequencing the heavy chain, light chain, variable region(s), and/orcomplementarity determining region(s) (CDR).

In addition to the enrichment step, the method for antibody selectioncan also include one or more steps of screening a cell population forantigen recognition and/or antibody functionality. For example, thedesired antibodies may have specific structural features, such asbinding to a particular epitope or mimicry of a particular structure;antagonist or agonist activity; or neutralizing activity, e.g.,inhibiting binding between the antigen and a ligand. In one embodiment,the antibody functionality screen is ligand-dependent. Screening forantibody functionality includes, but is not limited to, an in vitroprotein-protein interaction assay that recreates the natural interactionof the antigen ligand with recombinant receptor protein; and acell-based response that is ligand dependent and easily monitored (e.g.,proliferation response). In one embodiment, the method for antibodyselection includes a step of screening the cell population for antibodyfunctionality by measuring the inhibitory concentration (IC50). In oneembodiment, at least one of the isolated, antigen-specific cellsproduces an antibody having an IC50 of less than about 100, 50, 30, 25,10 μg/mL, or increments therein.

In addition to the enrichment step, the method for antibody selectioncan also include one or more steps of screening a cell population forantibody binding strength. Antibody binding strength can be measured byany method known in the art (e.g., Biacore). In one embodiment, at leastone of the isolated, antigen-specific cells produces an antibody havinga high antigen affinity, e.g., a dissociation constant (Kd) of less thanabout 5×10⁻¹⁰ M-1, preferably about 1×10⁻¹³ to 5×10⁻¹⁰, 1×10⁻¹² to1×10⁻¹⁰, 1×10⁻¹² to 7.5×10⁻¹¹, 1×10⁻¹¹ to 2×10⁻¹¹, about 1.5×10⁻¹¹ orless, or increments therein. In this embodiment, the antibodies are saidto be affinity mature. In a preferred embodiment, the affinity of theantibodies is comparable to or higher than the affinity of any one ofPanorex® (edrecolomab), Rituxan® (rituximab), Herceptin® (traztuzumab),Mylotarg® (gentuzumab), Campath® (alemtuzumab), Zevalin™ (ibritumomab),Erbitux™ (cetuximab), Avastin™ (bevicizumab), Raptiva™ (efalizumab),Remicade® (infliximab), Humira™ (adalimumab), and Xolair™ (omalizumab).Preferably, the affinity of the antibodies is comparable to or higherthan the affinity of Humira™. The affinity of an antibody can also beincreased by known affinity maturation techniques. In one embodiment, atleast one cell population is screened for at least one of, preferablyboth, antibody functionality and antibody binding strength.

In addition to the enrichment step, the method for antibody selectioncan also include one or more steps of screening a cell population forantibody sequence homology, especially human homology. In oneembodiment, at least one of the isolated, antigen-specific cellsproduces an antibody that has a homology to a human antibody of about50% to about 100%, or increments therein, or greater than about 60%,70%, 80%, 85%, 90%, or 95% homologous. The antibodies can be humanizedto increase the homology to a human sequence by techniques known in theart such as CDR grafting or selectivity determining residue grafting(SDR).

In another embodiment, the present invention also provides theantibodies themselves according to any of the embodiments describedabove in terms of IC50, Kd, and/or homology.

The B cell selection protocol disclosed herein has a number of intrinsicadvantages versus other methods for obtaining antibody-secreting B cellsand monoclonal antibodies specific to desired target antigens. Theseadvantages include, but are not restricted to, the following:

First, it has been found that when these selection procedures areutilized with a desired antigen such as IL-6 or TNF-α, the methodsreproducibly result in antigen-specific B cells capable of generatingwhat appears to be a substantially comprehensive complement ofantibodies, i.e., antibodies that bind to the various different epitopesof the antigen. Without being bound by theory, it is hypothesized thatthe comprehensive complement is attributable to the antigen enrichmentstep that is performed prior to initial B cell recovery. Moreover, thisadvantage allows for the isolation and selection of antibodies withdifferent properties as these properties may vary depending on theepitopic specificity of the particular antibody.

Second, it has been found that the B cell selection protocolreproducibly yields a clonal B cell culture containing a single B cell,or its progeny, secreting a single monoclonal antibody that generallybinds to the desired antigen with a relatively high binding affinity,i.e. picomolar or better antigen binding affinities. By contrast, priorantibody selection methods tend to yield relatively few high affinityantibodies and therefore require extensive screening procedures toisolate an antibody with therapeutic potential. Without being bound bytheory, it is hypothesized that the protocol results in both in vivo Bcell immunization of the host (primary immunization) followed by asecond in vitro B cell stimulation (secondary antigen priming step) thatmay enhance the ability and propensity of the recovered clonal B cellsto secrete a single high affinity monoclonal antibody specific to theantigen target.

Third, it has been observed (as shown herein with IL-6 specific B cells)that the B cell selection protocol reproducibly yields enriched B cellsproducing IgG's that are, on average, highly selective (antigenspecific) to the desired target. Antigen-enriched B cells recovered bythese methods are believed to contain B cells capable of yielding thedesired full complement of epitopic specificities as discussed above.

Fourth, it has been observed that the B cell selection protocols, evenwhen used with small antigens, i.e., peptides of 100 amino acids orless, e.g., 5-50 amino acids long, reproducibly give rise to a clonal Bcell culture that secretes a single high affinity antibody to the smallantigen, e.g., a peptide. This is highly surprising as it is generallyquite difficult, labor intensive, and sometimes not even feasible toproduce high affinity antibodies to small peptides. Accordingly, theinvention can be used to produce therapeutic antibodies to desiredpeptide targets, e.g., viral, bacterial or autoantigen peptides, therebyallowing for the production of monoclonal antibodies with very discretebinding properties or even the production of a cocktail of monoclonalantibodies to different peptide targets, e.g., different viral strains.This advantage may especially be useful in the context of the productionof a therapeutic or prophylactic vaccine having a desired valency, suchas an HPV vaccine that induces protective immunity to different HPVstrains.

Fifth, the B cell selection protocol, particularly when used with Bcells derived from rabbits, tends to reproducibly yield antigen-specificantibody sequences that are very similar to endogenous humanimmunoglobulins (around 90% similar at the amino acid level) and thatcontain CDRs that possess a length very analogous to humanimmunoglobulins and therefore require little or no sequence modification(typically at most only a few CDR residues may be modified in the parentantibody sequence and no framework exogenous residues introduced) inorder to eliminate potential immunogenicity concerns. In particular,preferably the recombinant antibody will contain only the host (rabbit)CDR1 and CDR2 residues required for antigen recognition and the entireCDR3. Thereby, the high antigen binding affinity of the recoveredantibody sequences produced according to the B cell and antibodyselection protocol remains intact or substantially intact even withhumanization.

In sum, these method can be used to produce antibodies exhibiting higherbinding affinities to more distinct epitopes by the use of a moreefficient protocol than was previously known.

In a specific embodiment, the present invention provides a method foridentifying a single B cell that secretes an antibody specific to adesired antigen and that optionally possesses at least one desiredfunctional property such as affinity, avidity, cytolytic activity, andthe like by a process including the following steps:

a. immunizing a host against an antigen;

b. harvesting B cells from the host;

c. enriching the harvested B cells to increase the frequency ofantigen-specific cells;

d. creating at least one single cell suspension;

e. culturing a sub-population from the single cell suspension underconditions that favor the survival of a single antigen-specific B cellper culture well;

f. isolating B cells from the sub-population; and

g. determining whether the single B cell produces an antibody specificto the antigen.

Typically, these methods will further comprise an additional step ofisolating and sequencing, in whole or in part, the polypeptide andnucleic acid sequences encoding the desired antibody. These sequences ormodified versions or portions thereof can be expressed in desired hostcells in order to produce recombinant antibodies to a desired antigen.

As noted previously, it is believed that the clonal population of Bcells predominantly comprises antibody-secreting B cells producingantibody against the desired antigen. It is also believed based onexperimental results obtained with several antigens and with different Bcell populations that the clonally produced B cells and the isolatedantigen-specific B cells derived therefrom produced according to theinvention secrete a monoclonal antibody that is typically of relativelyhigh affinity and moreover is capable of efficiently and reproduciblyproducing a selection of monoclonal antibodies of greater epitopicvariability as compared to other methods of deriving monoclonalantibodies from cultured antigen-specific B cells. In an exemplaryembodiment the population of immune cells used in such B cell selectionmethods will be derived from a rabbit. However, other hosts that produceantibodies, including non-human and human hosts, can alternatively beused as a source of immune B cells. It is believed that the use ofrabbits as a source of B cells may enhance the diversity of monoclonalantibodies that may be derived by the methods. Also, the antibodysequences derived from rabbits according to the invention typicallypossess sequences having a high degree of sequence identity to humanantibody sequences making them favored for use in humans since theyshould possess little antigenicity. In the course of humanization, thefinal humanized antibody contains a much lower foreign/host residuecontent, usually restricted to a subset of the host CDR residues thatdiffer dramatically due to their nature versus the human target sequenceused in the grafting. This enhances the probability of complete activityrecovery in the humanized antibody protein.

The methods of antibody selection using an enrichment step disclosedherein include a step of obtaining a immune cell-containing cellpopulation from an immunized host. Methods of obtaining an immunecell-containing cell population from an immunized host are known in theart and generally include inducing an immune response in a host andharvesting cells from the host to obtain one or more cell populations.The response can be elicited by immunizing the host against a desiredantigen. Alternatively, the host used as a source of such immune cellscan be naturally exposed to the desired antigen such as an individualwho has been infected with a particular pathogen such as a bacterium orvirus or alternatively has mounted a specific antibody response to acancer that the individual is afflicted with.

Host animals are well-known in the art and include, but are not limitedto, guinea pig, rabbit, mouse, rat, non-human primate, human, as well asother mammals and rodents, chicken, cow, pig, goat, and sheep.Preferably the host is a mammal, more preferably, rabbit, mouse, rat, orhuman. When exposed to an antigen, the host produces antibodies as partof the native immune response to the antigen. As mentioned, the immuneresponse can occur naturally, as a result of disease, or it can beinduced by immunization with the antigen. Immunization can be performedby any method known in the art, such as, by one or more injections ofthe antigen with or without an agent to enhance immune response, such ascomplete or incomplete Freund's adjuvant. In another embodiment, theinvention also contemplates intrasplenic immunization. As an alternativeto immunizing a host animal in vivo, the method can comprise immunizinga host cell culture in vitro.

After allowing time for the immune response (e.g., as measured by serumantibody detection), host animal cells are harvested to obtain one ormore cell populations. In a preferred embodiment, a harvested cellpopulation is screened for antibody binding strength and/or antibodyfunctionality. A harvested cell population is preferably from at leastone of the spleen, lymph nodes, bone marrow, and/or peripheral bloodmononuclear cells (PBMCs). The cells can be harvested from more than onesource and pooled. Certain sources may be preferred for certainantigens. For example, the spleen, lymph nodes, and PBMCs are preferredfor IL-6; and the lymph nodes are preferred for TNF. The cell populationis harvested about 20 to about 90 days or increments therein afterimmunization, preferably about 50 to about 60 days. A harvested cellpopulation and/or a single cell suspension therefrom can be enriched,screened, and/or cultured for antibody selection. The frequency ofantigen-specific cells within a harvested cell population is usuallyabout 1% to about 5%, or increments therein.

In one embodiment, a single cell suspension from a harvested cellpopulation is enriched, preferably by using Miltenyi beads. From theharvested cell population having a frequency of antigen-specific cellsof about 1% to about 5%, an enriched cell population is thus derivedhaving a frequency of antigen-specific cells approaching 100%.

The method of antibody selection using an enrichment step includes astep of producing antibodies from at least one antigen-specific cellfrom an enriched cell population. Methods of producing antibodies invitro are well known in the art, and any suitable method can beemployed. In one embodiment, an enriched cell population, such as anantigen-specific single cell suspension from a harvested cellpopulation, is plated at various cell densities, such as 50, 100, 250,500, or other increments between 1 and 1000 cells per well. Preferably,the sub-population comprises no more than about 10,000 antigen-specific,antibody-secreting cells, more preferably about 50-10,000, about50-5,000, about 50-1,000, about 50-500, about 50-250 antigen-specific,antibody-secreting cells, or increments therein. Then, thesesub-populations are cultured with suitable medium (e.g., an activated Tcell conditioned medium, particularly 1-5% activated rabbit T cellconditioned medium) on a feeder layer, preferably under conditions thatfavor the survival of a single proliferating antibody-secreting cell perculture well. The feeder layer, generally comprised of irradiated cellmatter, e.g., EL4B cells, does not constitute part of the cellpopulation. The cells are cultured in a suitable media for a timesufficient for antibody production, for example about 1 day to about 2weeks, about 1 day to about 10 days, at least about 3 days, about 3 toabout 5 days, about 5 days to about 7 days, at least about 7 days, orother increments therein. In one embodiment, more than onesub-population is cultured simultaneously. Preferably, a singleantibody-producing cell and progeny thereof survives in each well,thereby providing a clonal population of antigen-specific B cells ineach well. At this stage, the immunoglobulin G (IgG) produced by theclonal population is highly correlative with antigen specificity. In apreferred embodiment, the IgGs exhibit a correlation with antigenspecificity that is greater than about 50%, more preferably greater than70%, 85%, 90%, 95%, 99%, or increments therein. See FIG. 3, whichdemonstrates an exemplary correlation for IL-6. The correlations weredemonstrated by setting up B cell cultures under limiting conditions toestablish single antigen-specific antibody products per well.Antigen-specific versus general IgG synthesis was compared. Threepopulations were observed: IgG that recognized a single format ofantigen (biotinylated and direct coating), detectable IgG and antigenrecognition irrespective of immobilization, and IgG production alone.IgG production was highly correlated with antigen-specificity.

A supernatant containing the antibodies is optionally collected, whichcan be can be enriched, screened, and/or cultured for antibody selectionaccording to the steps described above. In one embodiment, thesupernatant is enriched (preferably by an antigen-specificity assay,especially an ELISA assay) and/or screened for antibody functionality.

In another embodiment, the enriched, preferably clonal, antigen-specificB cell population from which a supernatant described above is optionallyscreened in order to detect the presence of the desired secretedmonoclonal antibody is used for the isolation of a few B cells,preferably a single B cell, which is then tested in an appropriate assayin order to confirm the presence of a single antibody-producing B cellin the clonal B cell population. In one embodiment about 1 to about 20cells are isolated from the clonal B cell population, preferably lessthan about 15, 12, 10, 5, or 3 cells, or increments therein, mostpreferably a single cell. The screen is preferably effected by anantigen-specificity assay, especially a halo assay. The halo assay canbe performed with the full length protein, or a fragment thereof. Theantibody-containing supernatant can also be screened for at least oneof: antigen binding affinity; agonism or antagonism of antigen-ligandbinding, induction or inhibition of the proliferation of a specifictarget cell type; induction or inhibition of lysis of a target cell, andinduction or inhibition of a biological pathway involving the antigen.

The identified antigen-specific cell can be used to derive thecorresponding nucleic acid sequences encoding the desired monoclonalantibody. (An AluI digest can confirm that only a single monoclonalantibody type is produced per well.) As mentioned above, these sequencescan be mutated, such as by humanization, in order to render themsuitable for use in human medicaments.

As mentioned, the enriched B cell population used in the process canalso be further enriched, screened, and/or cultured for antibodyselection according to the steps described above which can be repeatedor performed in a different order. In a preferred embodiment, at leastone cell of an enriched, preferably clonal, antigen-specific cellpopulation is isolated, cultured, and used for antibody selection.

Thus, in one embodiment, the present invention provides a methodcomprising:

a. harvesting a cell population from an immunized host to obtain aharvested cell population;

b. creating at least one single cell suspension from a harvested cellpopulation;

c. enriching at least one single cell suspension, preferably bychromatography, to form a first enriched cell population;

d. enriching the first enriched cell population, preferably by ELISAassay, to form a second enriched cell population which preferably isclonal, i.e., it contains only a single type of antigen-specific B cell;

e. enriching the second enriched cell population, preferably by haloassay, to form a third enriched cell population containing a single or afew number of B cells that produce an antibody specific to a desiredantigen; and

f. selecting an antibody produced by an antigen-specific cell isolatedfrom the third enriched cell population.

The method can further include one or more steps of screening theharvested cell population for antibody binding strength (affinity,avidity) and/or antibody functionality. Suitable screening stepsinclude, but are not limited to, assay methods that detect: whether theantibody produced by the identified antigen-specific B cell produces anantibody possessing a minimal antigen binding affinity, whether theantibody agonizes or antagonizes the binding of a desired antigen to aligand; whether the antibody induces or inhibits the proliferation of aspecific cell type; whether the antibody induces or elicits a cytolyticreaction against target cells; whether the antibody binds to a specificepitope; and whether the antibody modulates (inhibits or agonizes) aspecific biological pathway or pathways involving the antigen.

Similarly, the method can include one or more steps of screening thesecond enriched cell population for antibody binding strength and/orantibody functionality.

The method can further include a step of sequencing the polypeptidesequence or the corresponding nucleic acid sequence of the selectedantibody. The method can also include a step of producing a recombinantantibody using the sequence, a fragment thereof, or a geneticallymodified version of the selected antibody. Methods for mutating antibodysequences in order to retain desired properties are well known to thoseskilled in the art and include humanization, chimerisation, productionof single chain antibodies; these mutation methods can yield recombinantantibodies possessing desired effector function, immunogenicity,stability, removal or addition of glycosylation, and the like. Therecombinant antibody can be produced by any suitable recombinant cell,including, but not limited to mammalian cells such as CHO, COS, BHK,HEK-293, bacterial cells, yeast cells, plant cells, insect cells, andamphibian cells. In one embodiment, the antibodies are expressed inpolyploidal yeast cells, i.e., diploid yeast cells, particularly Pichia.

In one embodiment, the method comprises:

a. immunizing a host against an antigen to yield host antibodies;

b. screening the host antibodies for antigen specificity andneutralization;

c. harvesting B cells from the host;

d. enriching the harvested B cells to create an enriched cell populationhaving an increased frequency of antigen-specific cells;

e. culturing one or more sub-populations from the enriched cellpopulation under conditions that favor the survival of a single B cellto produce a clonal population in at least one culture well;

f. determining whether the clonal population produces an antibodyspecific to the antigen;

g. isolating a single B cell; and

h. sequencing the nucleic acid sequence of the antibody produced by thesingle B cell.

Methods of Humanizing Antibodies

In another embodiment of the invention, there is provided a method forhumanizing antibody heavy and light chains. In this embodiment, thefollowing method is followed for the humanization of the heavy and lightchains:

Light Chain

1. Identify the amino acid that is the first one following the signalpeptide sequence. This is the start of Framework 1. The signal peptidestarts at the first initiation methionine and is typically, but notnecessarily 22 amino acids in length for rabbit light chain proteinsequences. The start of the mature polypeptide can also be determinedexperimentally by N-terminal protein sequencing, or can be predictedusing a prediction algorithm. This is also the start of Framework 1 asclassically defined by those in the field.

Example: RbtVL Amino acid residue 1 in FIG. 2, starting ‘AYDM . . . ’

2. Identify the end of Framework 3. This is typically 86-90 amino acidsfollowing the start of Framework 1 and is typically a cysteine residuepreceded by two tyrosine residues. This is the end of the Framework 3 asclassically defined by those in the field.

Example: RbtVL amino acid residue 88 in FIG. 2, ending as ‘TYYC’

3. Use the rabbit light chain sequence of the polypeptide starting fromthe beginning of Framework 1 to the end of Framework 3 as defined aboveand perform a sequence homology search for the most similar humanantibody protein sequences. This will typically be a search againsthuman germline sequences prior to antibody maturation in order to reducethe possibility of immunogenicity, however any human sequences can beused. Typically a program like BLAST can be used to search a database ofsequences for the most homologous. Databases of human antibody sequencescan be found from various sources such as NCBI (National Center forBiotechnology Information).

Example: RbtVL amino acid sequence from residues numbered 1 through 88in FIG. 2 is BLASTed against a human antibody germline database. The topthree unique returned sequences are shown in FIG. 2 as L12A, V1 andVx02.

4. Generally the most homologous human germline variable light chainsequence is then used as the basis for humanization. However thoseskilled in the art may decide to use another sequence that wasn't thehighest homology as determined by the homology algorithm, based on otherfactors including sequence gaps and framework similarities.

Example: In FIG. 2, L12A was the most homologous human germline variablelight chain sequence and is used as the basis for the humanization ofRbtVL.

5. Determine the framework and CDR arrangement (FR1, FR2, FR3, CDR1 &CDR2) for the human homolog being used for the light chain humanization.This is using the traditional layout as described in the field. Alignthe rabbit variable light chain sequence with the human homolog, whilemaintaining the layout of the framework and CDR regions.

Example: In FIG. 2, the RbtVL sequence is aligned with the humanhomologous sequence L12A, and the framework and CDR domains areindicated.

6. Replace the human homologous light chain sequence CDR1 and CDR2regions with the CDR1 and CDR2 sequences from the rabbit sequence. Ifthere are differences in length between the rabbit and human CDRsequences then use the entire rabbit CDR sequences and their lengths. Itis possible that the specificity, affinity and/or immunogenicity of theresulting humanized antibody may be unaltered if smaller or largersequence exchanges are performed, or if specific residue(s) are altered,however the exchanges as described have been used successfully, but donot exclude the possibility that other changes may be permitted.

Example: In FIG. 2, the CDR1 and CDR2 amino acid residues of the humanhomologous variable light chain L12A are replaced with the CDR1 and CDR2amino acid sequences from the RbtVL rabbit antibody light chainsequence. The human L12A frameworks 1, 2 and 3 are unaltered. Theresulting humanized sequence is shown below as VLh from residuesnumbered 1 through 88. Note that the only residues that are differentfrom the L12A human sequence are underlined, and are thus rabbit-derivedamino acid residues. In this example only 8 of the 88 residues aredifferent than the human sequence.

7. After framework 3 of the new hybrid sequence created in Step 6,attach the entire CDR3 of the rabbit light chain antibody sequence. TheCDR3 sequence can be of various lengths, but is typically 9 to 15 aminoacid residues in length. The CDR3 region and the beginning of thefollowing framework 4 region are defined classically and identifiable bythose skilled in the art. Typically the beginning of Framework 4, andthus after the end of CDR3 consists of the sequence ‘FGGG . . . ’ (SEQID NO:666), however some variation may exist in these residues.

Example: In FIG. 2, the CDR3 of RbtVL (amino acid residues numbered89-100) is added after the end of framework 3 in the humanized sequenceindicated as VLh.

8. The rabbit light chain framework 4, which is typically the final 11amino acid residues of the variable light chain and begins as indicatedin Step 7 above and typically ends with the amino acid sequence ‘ . . .VVKR’ (SEQ ID NO:667) is replaced with the nearest human light chainframework 4 homolog, usually from germline sequence. Frequently thishuman light chain framework 4 is of the sequence ‘FGGGTKVEIKR’ (SEQ IDNO:668). It is possible that other human light chain framework 4sequences that are not the most homologous or otherwise different may beused without affecting the specificity, affinity and/or immunogenicityof the resulting humanized antibody. This human light chain framework 4sequence is added to the end of the variable light chain humanizedsequence immediately following the CDR3 sequence from Step 7 above. Thisis now the end of the variable light chain humanized amino acidsequence.

Example: In FIG. 2, Framework 4 (FR4) of the RbtVL rabbit light chainsequence is shown above a homologous human FR4 sequence. The human FR4sequence is added to the humanized variable light chain sequence (VLh)right after the end of the CD3 region added in Step 7 above.

Heavy Chain

1. Identify the amino acid that is the first one following the signalpeptide sequence. This is the start of Framework 1. The signal peptidestarts at the first initiation methionine and is typically 19 aminoacids in length for rabbit heavy chain protein sequences. Typically, butnot necessarily always, the final 3 amino acid residues of a rabbitheavy chain signal peptide are ‘ . . . VQC’, followed by the start ofFramework 1. The start of the mature polypeptide can also be determinedexperimentally by N-terminal protein sequencing, or can be predictedusing a prediction algorithm. This is also the start of Framework 1 asclassically defined by those in the field.

Example: RbtVH Amino acid residue 1 in FIG. 2, starting ‘QEQL . . . ’

2. Identify the end of Framework 3. This is typically 95-100 amino acidsfollowing the start of Framework 1 and typically has the final sequenceof ‘ . . . CAR’ (although the alanine can also be a valine). This is theend of the Framework 3 as classically defined by those in the field.

Example: RbtVH amino acid residue 98 in FIG. 2, ending as ‘ . . . FCVR’.

3. Use the rabbit heavy chain sequence of the polypeptide starting fromthe beginning of Framework 1 to the end of Framework 3 as defined aboveand perform a sequence homology search for the most similar humanantibody protein sequences. This will typically be against a database ofhuman germline sequences prior to antibody maturation in order to reducethe possibility of immunogenicity, however any human sequences can beused. Typically a program like BLAST can be used to search a database ofsequences for the most homologous. Databases of human antibody sequencescan be found from various sources such as NCBI (National Center forBiotechnology Information).

Example: RbtVH amino acid sequence from residues numbered 1 through 98in FIG. 2 is BLASTed against a human antibody germline database. The topthree unique returned sequences are shown in FIG. 2 as 3-64-04, 3-66-04,and 3-53-02.

4. Generally the most homologous human germline variable heavy chainsequence is then used as the basis for humanization. However thoseskilled in the art may decide to use another sequence that wasn't themost homologous as determined by the homology algorithm, based on otherfactors including sequence gaps and framework similarities.

Example: 3-64-04 in FIG. 2 was the most homologous human germlinevariable heavy chain sequence and is used as the basis for thehumanization of RbtVH.

5. Determine the framework and CDR arrangement (FR1, FR2, FR3, CDR1 &CDR2) for the human homolog being used for the heavy chain humanization.This is using the traditional layout as described in the field. Alignthe rabbit variable heavy chain sequence with the human homolog, whilemaintaining the layout of the framework and CDR regions.

Example: In FIG. 2, the RbtVH sequence is aligned with the humanhomologous sequence 3-64-04, and the framework and CDR domains areindicated.

6. Replace the human homologous heavy chain sequence CDR1 and CDR2regions with the CDR1 and CDR2 sequences from the rabbit sequence. Ifthere are differences in length between the rabbit and human CDRsequences then use the entire rabbit CDR sequences and their lengths. Inaddition, it may be necessary to replace the final three amino acids ofthe human heavy chain Framework 1 region with the final three aminoacids of the rabbit heavy chain Framework 1. Typically but not always,in rabbit heavy chain Framework 1 these three residues follow a Glycineresidue preceded by a Serine residue. In addition, it may be necessaryreplace the final amino acid of the human heavy chain Framework 2 regionwith the final amino acid of the rabbit heavy chain Framework 2.Typically, but not necessarily always, this is a Glycine residuepreceded by an Isoleucine residue in the rabbit heavy chain Framework 2.It is possible that the specificity, affinity and/or immunogenicity ofthe resulting humanized antibody may be unaltered if smaller or largersequence exchanges are performed, or if specific residue(s) are altered,however the exchanges as described have been used successfully, but donot exclude the possibility that other changes may be permitted. Forexample, a tryptophan amino acid residue typically occurs four residuesprior to the end of the rabbit heavy chain CDR2 region, whereas in humanheavy chain CDR2 this residue is typically a Serine residue. Changingthis rabbit tryptophan residue to a the human Serine residue at thisposition has been demonstrated to have minimal to no effect on thehumanized antibody's specificity or affinity, and thus further minimizesthe content of rabbit sequence-derived amino acid residues in thehumanized sequence.

Example: In FIG. 2, The CDR1 and CDR2 amino acid residues of the humanhomologous variable heavy chain are replaced with the CDR1 and CDR2amino acid sequences from the RbtVH rabbit antibody light chainsequence, except for the boxed residue, which is tryptophan in therabbit sequence (position number 63) and Serine at the same position inthe human sequence, and is kept as the human Serine residue. In additionto the CDR1 and CDR2 changes, the final three amino acids of Framework 1(positions 28-30) as well as the final residue of Framework 2 (position49) are retained as rabbit amino acid residues instead of human. Theresulting humanized sequence is shown below as VHh from residuesnumbered 1 through 98. Note that the only residues that are differentfrom the 3-64-04 human sequence are underlined, and are thusrabbit-derived amino acid residues. In this example only 15 of the 98residues are different than the human sequence.

7. After framework 3 of the new hybrid sequence created in Step 6,attach the entire CDR3 of the rabbit heavy chain antibody sequence. TheCDR3 sequence can be of various lengths, but is typically 5 to 19 aminoacid residues in length. The CDR3 region and the beginning of thefollowing framework 4 region are defined classically and areidentifiable by those skilled in the art. Typically the beginning offramework 4, and thus after the end of CDR3 consists of the sequenceWGXG . . . (where X is usually Q or P), however some variation may existin these residues.

Example: The CDR3 of RbtVH (amino acid residues numbered 99-110) isadded after the end of framework 3 in the humanized sequence indicatedas VHh.

8. The rabbit heavy chain framework 4, which is typically the final 11amino acid residues of the variable heavy chain and begins as indicatedin Step 7 above and typically ends with the amino acid sequence ‘ . . .TVSS’ is replaced with the nearest human heavy chain framework 4homolog, usually from germline sequence. Frequently this human heavychain framework 4 is of the sequence ‘WGQGTLVTVSS’. It is possible thatother human heavy chain framework 4 sequences that are not the mosthomologous or otherwise different may be used without affecting thespecificity, affinity and/or immunogenicity of the resulting humanizedantibody. This human heavy chain framework 4 sequence is added to theend of the variable heavy chain humanized sequence immediately followingthe CDR3 sequence from Step 7 above. This is now the end of the variableheavy chain humanized amino acid sequence.

Example: In FIG. 2, framework 4 (FR4) of the RbtVH rabbit heavy chainsequence is shown above a homologous human heavy FR4 sequence. The humanFR4 sequence is added to the humanized variable heavy chain sequence(VHh) right after the end of the CD3 region added in Step 7 above.

Methods of Producing Antibodies and Fragments Thereof

The invention is also directed to the production of the antibodiesdescribed herein or fragments thereof. Recombinant polypeptidescorresponding to the antibodies described herein or fragments thereofare secreted from polyploidal, preferably diploid or tetraploid strainsof mating competent yeast. In an exemplary embodiment, the invention isdirected to methods for producing these recombinant polypeptides insecreted form for prolonged periods using cultures comprising polyploidyeast, i.e., at least several days to a week, more preferably at least amonth or several months, and even more preferably at least 6 months to ayear or longer. These polyploid yeast cultures will express at least10-25 mg/liter of the polypeptide, more preferably at least 50-250mg/liter, still more preferably at least 500-1000 mg/liter, and mostpreferably a gram per liter or more of the recombinant polypeptide(s).

In one embodiment of the invention a pair of genetically marked yeasthaploid cells are transformed with expression vectors comprisingsubunits of a desired heteromultimeric protein. One haploid cellcomprises a first expression vector, and a second haploid cell comprisesa second expression vector. In another embodiment diploid yeast cellswill be transformed with one or more expression vectors that provide forthe expression and secretion of one or more of the recombinantpolypeptides. In still another embodiment a single haploid cell may betransformed with one or more vectors and used to produce a polyploidalyeast by fusion or mating strategies. In yet another embodiment adiploid yeast culture may be transformed with one or more vectorsproviding for the expression and secretion of a desired polypeptide orpolypeptides. These vectors may comprise vectors e.g., linearizedplasmids or other linear DNA products that integrate into the yeastcell's genome randomly, through homologous recombination, or using arecombinase such as Cre/Lox or Flp/Frt. Optionally, additionalexpression vectors may be introduced into the haploid or diploid cells;or the first or second expression vectors may comprise additional codingsequences; for the synthesis of heterotrimers; heterotetramers; etc. Theexpression levels of the non-identical polypeptides may be individuallycalibrated, and adjusted through appropriate selection, vector copynumber, promoter strength and/or induction and the like. The transformedhaploid cells are genetically crossed or fused. The resulting diploid ortetraploid strains are utilized to produce and secrete fully assembledand biologically functional proteins, humanized antibodies describedherein or fragments thereof.

The use of diploid or tetraploid cells for protein production providesfor unexpected benefits. The cells can be grown for production purposes,i.e. scaled up, and for extended periods of time, in conditions that canbe deleterious to the growth of haploid cells, which conditions mayinclude high cell density; growth in minimal media; growth at lowtemperatures; stable growth in the absence of selective pressure; andwhich may provide for maintenance of heterologous gene sequenceintegrity and maintenance of high level expression over time. Withoutwishing to be bound thereby, the inventors theorize that these benefitsmay arise, at least in part, from the creation of diploid strains fromtwo distinct parental haploid strains. Such haploid strains can comprisenumerous minor autotrophic mutations, which mutations are complementedin the diploid or tetraploid, enabling growth and enhanced productionunder highly selective conditions.

Transformed mating competent haploid yeast cells provide a geneticmethod that enables subunit pairing of a desired protein. Haploid yeaststrains are transformed with each of two expression vectors, a firstvector to direct the synthesis of one polypeptide chain and a secondvector to direct the synthesis of a second, non-identical polypeptidechain. The two haploid strains are mated to provide a diploid host whereoptimized target protein production can be obtained.

Optionally, additional non-identical coding sequence(s) are provided.Such sequences may be present on additional expression vectors or in thefirst or the second expression vectors. As is known in the art, multiplecoding sequences may be independently expressed from individualpromoters; or may be coordinately expressed through the inclusion of an“internal ribosome entry site” or “IRES”, which is an element thatpromotes direct internal ribosome entry to the initiation codon, such asATG, of a cistron (a protein encoding region), thereby leading to thecap-independent translation of the gene. IRES elements functional inyeast are described by Thompson et al. (2001) P.N.A.S. 98:12866-12868.

In one embodiment of the invention, antibody sequences are produced incombination with a secretory J chain, which provides for enhancedstability of IgA (see U.S. Pat. Nos. 5,959,177; and 5,202,422).

In a preferred embodiment the two haploid yeast strains are eachauxotrophic, and require supplementation of media for growth of thehaploid cells. The pair of auxotrophs are complementary, such that thediploid product will grow in the absence of the supplements required forthe haploid cells. Many such genetic markers are known in yeast,including requirements for amino acids (e.g. met, lys, his, arg, etc.),nucleosides (e.g. ura3, ade1, etc.); and the like. Amino acid markersmay be preferred for the methods of the invention. Alternatively diploidcells which contain the desired vectors can be selected by other means,e.g., by use of other markers, such as green fluorescent protein,antibiotic resistance genes, various dominant selectable markers, andthe like.

Two transformed haploid cells may be genetically crossed and diploidstrains arising from this mating event selected by their hybridnutritional requirements and/or antibiotic resistance spectra.Alternatively, populations of the two transformed haploid strains arespheroplasted and fused, and diploid progeny regenerated and selected.By either method, diploid strains can be identified and selectivelygrown based on their ability to grow in different media than theirparents. For example, the diploid cells may be grown in minimal mediumthat may include antibiotics. The diploid synthesis strategy has certainadvantages. Diploid strains have the potential to produce enhancedlevels of heterologous protein through broader complementation tounderlying mutations, which may impact the production and/or secretionof recombinant protein. Furthermore, once stable strains have beenobtained, any antibiotics used to select those strains do notnecessarily need to be continuously present in the growth media.

As noted above, in some embodiments a haploid yeast may be transformedwith a single or multiple vectors and mated or fused with anon-transformed cell to produce a diploid cell containing the vector orvectors. In other embodiments, a diploid yeast cell may be transformedwith one or more vectors that provide for the expression and secretionof a desired heterologous polypeptide by the diploid yeast cell.

In one embodiment of the invention, two haploid strains are transformedwith a library of polypeptides, e.g. a library of antibody heavy orlight chains. Transformed haploid cells that synthesize the polypeptidesare mated with the complementary haploid cells. The resulting diploidcells are screened for functional protein. The diploid cells provide ameans of rapidly, conveniently and inexpensively bringing together alarge number of combinations of polypeptides for functional testing.This technology is especially applicable for the generation ofheterodimeric protein products, where optimized subunit synthesis levelsare critical for functional protein expression and secretion.

In another embodiment of the invention, the expression level ratio ofthe two subunits is regulated in order to maximize product generation.Heterodimer subunit protein levels have been shown previously to impactthe final product generation (Simmons L C, J Immunol Methods. 2002 May1; 263(1-2):133-47). Regulation can be achieved prior to the mating stepby selection for a marker present on the expression vector. By stablyincreasing the copy number of the vector, the expression level can beincreased. In some cases, it may be desirable to increase the level ofone chain relative to the other, so as to reach a balanced proportionbetween the subunits of the polypeptide. Antibiotic resistance markersare useful for this purpose, e.g. Zeocin resistance marker, G418resistance, etc. and provide a means of enrichment for strains thatcontain multiple integrated copies of an expression vector in a strainby selecting for transformants that are resistant to higher levels ofZeocin or G418. The proper ratio, e.g. 1:1; 1:2; etc. of the subunitgenes may be important for efficient protein production. Even when thesame promoter is used to transcribe both subunits, many other factorscontribute to the final level of protein expressed and therefore, it canbe useful to increase the number of copies of one encoded gene relativeto the other. Alternatively, diploid strains that produce higher levelsof a polypeptide, relative to single copy vector strains, are created bymating two haploid strains, both of which have multiple copies of theexpression vectors.

Host cells are transformed with the above-described expression vectors,mated to form diploid strains, and cultured in conventional nutrientmedia modified as appropriate for inducing promoters, selectingtransformants or amplifying the genes encoding the desired sequences. Anumber of minimal media suitable for the growth of yeast are known inthe art. Any of these media may be supplemented as necessary with salts(such as sodium chloride, calcium, magnesium, and phosphate), buffers(such as phosphate, HEPES), nucleosides (such as adenosine andthymidine), antibiotics, trace elements, and glucose or an equivalentenergy source. Any other necessary supplements may also be included atappropriate concentrations that would be known to those skilled in theart. The culture conditions, such as temperature, pH and the like, arethose previously used with the host cell selected for expression, andwill be apparent to the ordinarily skilled artisan.

Secreted proteins are recovered from the culture medium. A proteaseinhibitor, such as phenyl methyl sulfonyl fluoride (PMSF) may be usefulto inhibit proteolytic degradation during purification, and antibioticsmay be included to prevent the growth of adventitious contaminants. Thecomposition may be concentrated, filtered, dialyzed, etc., using methodsknown in the art.

The diploid cells of the invention are grown for production purposes.Such production purposes desirably include growth in minimal media,which media lacks pre-formed amino acids and other complex biomolecules,e.g., media comprising ammonia as a nitrogen source, and glucose as anenergy and carbon source, and salts as a source of phosphate, calciumand the like. Preferably such production media lacks selective agentssuch as antibiotics, amino acids, purines, pyrimidines, etc. The diploidcells can be grown to high cell density, for example at least about 50g/L; more usually at least about 100 g/L; and may be at least about 300,about 400, about 500 g/L or more.

In one embodiment of the invention, the growth of the subject cells forproduction purposes is performed at low temperatures, which temperaturesmay be lowered during log phase, during stationary phase, or both. Theterm “low temperature” refers to temperatures of at least about 15° C.,more usually at least about 17° C., and may be about 20° C., and isusually not more than about 25° C., more usually not more than about 22°C. In another embodiment of the invention, the low temperature isusually not more than about 28° C. Growth temperature can impact theproduction of full-length secreted proteins in production cultures, anddecreasing the culture growth temperature can strongly enhance theintact product yield. The decreased temperature appears to assistintracellular trafficking through the folding and post-translationalprocessing pathways used by the host to generate the target product,along with reduction of cellular protease degradation.

The methods of the invention provide for expression of secreted, activeprotein, preferably a mammalian protein. In one embodiment, secreted,“active antibodies”, as used herein, refers to a correctly foldedmultimer of at least two properly paired chains, which accurately bindsto its cognate antigen. Expression levels of active protein are usuallyat least about 10-50 mg/liter culture, more usually at least about 100mg/liter, preferably at least about 500 mg/liter, and may be 1000mg/liter or more.

The methods of the invention can provide for increased stability of thehost and heterologous coding sequences during production. The stabilityis evidenced, for example, by maintenance of high levels of expressionof time, where the starting level of expression is decreased by not morethan about 20%, usually not more than 10%, and may be decreased by notmore than about 5% over about 20 doublings, 50 doublings, 100 doublings,or more.

The strain stability also provides for maintenance of heterologous genesequence integrity over time, where the sequence of the active codingsequence and requisite transcriptional regulatory elements aremaintained in at least about 99% of the diploid cells, usually in atleast about 99.9% of the diploid cells, and preferably in at least about99.99% of the diploid cells over about 20 doublings, 50 doublings, 100doublings, or more. Preferably, substantially all of the diploid cellsmaintain the sequence of the active coding sequence and requisitetranscriptional regulatory elements.

Other methods of producing antibodies are well known to those ofordinary skill in the art. For example, methods of producing chimericantibodies are now well known in the art (See, for example, U.S. Pat.No. 4,816,567 to Cabilly et al.; Morrison et al., P.N.A.S. USA,81:8651-55 (1984); Neuberger, M. S. et al., Nature, 314:268-270 (1985);Boulianne, G. L. et al., Nature, 312:643-46 (1984), the disclosures ofeach of which are herein incorporated by reference in their entireties).

Likewise, other methods of producing humanized antibodies are now wellknown in the art (See, for example, U.S. Pat. Nos. 5,530,101, 5,585,089,5,693,762, and 6,180,370 to Queen et al; U.S. Pat. Nos. 5,225,539 and6,548,640 to Winter; U.S. Pat. Nos. 6,054,297, 6,407,213 and 6,639,055to Carter et al; U.S. Pat. No. 6,632,927 to Adair; Jones, P. T. et al,Nature, 321:522-525 (1986); Reichmann, L., et al, Nature, 332:323-327(1988); Verhoeyen, M, et al, Science, 239:1534-36 (1988), thedisclosures of each of which are herein incorporated by reference intheir entireties).

Antibody polypeptides of the invention having IL-6 binding specificitymay also be produced by constructing, using conventional techniques wellknown to those of ordinary skill in the art, an expression vectorcontaining an operon and a DNA sequence encoding an antibody heavy chainin which the DNA sequence encoding the CDRs required for antibodyspecificity is derived from a non-human cell source, preferably a rabbitB-cell source, while the DNA sequence encoding the remaining parts ofthe antibody chain is derived from a human cell source.

A second expression vector is produced using the same conventional meanswell known to those of ordinary skill in the art, said expression vectorcontaining an operon and a DNA sequence encoding an antibody light chainin which the DNA sequence encoding the CDRs required for antibodyspecificity is derived from a non-human cell source, preferably a rabbitB-cell source, while the DNA sequence encoding the remaining parts ofthe antibody chain is derived from a human cell source.

The expression vectors are transfected into a host cell by conventiontechniques well known to those of ordinary skill in the art to produce atransfected host cell, said transfected host cell cultured byconventional techniques well known to those of ordinary skill in the artto produce said antibody polypeptides.

The host cell may be co-transfected with the two expression vectorsdescribed above, the first expression vector containing DNA encoding anoperon and a light chain-derived polypeptide and the second vectorcontaining DNA encoding an operon and a heavy chain-derived polypeptide.The two vectors contain different selectable markers, but preferablyachieve substantially equal expression of the heavy and light chainpolypeptides. Alternatively, a single vector may be used, the vectorincluding DNA encoding both the heavy and light chain polypeptides. Thecoding sequences for the heavy and light chains may comprise cDNA.

The host cells used to express the antibody polypeptides may be either abacterial cell such as E. coli, or a eukaryotic cell. In a particularlypreferred embodiment of the invention, a mammalian cell of awell-defined type for this purpose, such as a myeloma cell or a Chinesehamster ovary (CHO) cell line may be used.

The general methods by which the vectors may be constructed,transfection methods required to produce the host cell and culturingmethods required to produce the antibody polypeptides from said hostcells all include conventional techniques. Although preferably the cellline used to produce the antibody is a mammalian cell line, any othersuitable cell line, such as a bacterial cell line such as an E.coli-derived bacterial strain, or a yeast cell line, may alternativelybe used.

Similarly, once produced the antibody polypeptides may be purifiedaccording to standard procedures in the art, such as for examplecross-flow filtration, ammonium sulphate precipitation, affinity columnchromatography and the like.

The antibody polypeptides described herein may also be used for thedesign and synthesis of either peptide or non-peptide mimetics thatwould be useful for the same therapeutic applications as the antibodypolypeptides of the invention. See, for example, Saragobi et al,Science, 253:792-795 (1991), the contents of which is hereinincorporated by reference in its entirety.

Screening Assays

The invention also includes screening assays designed to assist in theidentification of diseases and disorders associated with IL-6 inpatients exhibiting symptoms of an IL-6 associated disease or disorder.

In one embodiment of the invention, the anti-IL-6 antibodies of theinvention, or IL-6 binding fragments thereof, are used to detect thepresence of IL-6 in a biological sample obtained from a patientexhibiting symptoms of a disease or disorder associated with IL-6. Thepresence of IL-6, or elevated levels thereof when compared topre-disease levels of IL-6 in a comparable biological sample, may bebeneficial in diagnosing a disease or disorder associated with IL-6.

Another embodiment of the invention provides a diagnostic or screeningassay to assist in diagnosis of diseases or disorders associated withIL-6 in patients exhibiting symptoms of an IL-6 associated disease ordisorder identified herein, comprising assaying the level of IL-6expression in a biological sample from said patient using apost-translationally modified anti-IL-6 antibody or binding fragmentthereof. The anti-IL-6 antibody or binding fragment thereof may bepost-translationally modified to include a detectable moiety such as setforth previously in the disclosure.

The IL-6 level in the biological sample is determined using a modifiedanti-IL-6 antibody or binding fragment thereof as set forth herein, andcomparing the level of IL-6 in the biological sample against a standardlevel of IL-6 (e.g., the level in normal biological samples). Theskilled clinician would understand that some variability may existbetween normal biological samples, and would take that intoconsideration when evaluating results.

The above-recited assay may also be useful in monitoring a disease ordisorder, where the level of IL-6 obtained in a biological sample from apatient believed to have an IL-6 associated disease or disorder iscompared with the level of IL-6 in prior biological samples from thesame patient, in order to ascertain whether the IL-6 level in saidpatient has changed with, for example, a treatment regimen.

The invention is also directed to a method of in vivo imaging whichdetects the presence of cells which express IL-6 comprisingadministering a diagnostically effective amount of a diagnosticcomposition. Said in vivo imaging is useful for the detection andimaging of IL-6 expressing tumors or metastases and IL-6 expressinginflammatory sites, for example, and can be used as part of a planningregimen for design of an effective cancer or arthritis treatmentprotocol. The treatment protocol may include, for example, one or moreof radiation, chemotherapy, cytokine therapy, gene therapy, and antibodytherapy, as well as an anti-IL-6 antibody or fragment thereof.

A skilled clinician would understand that a biological sample includes,but is not limited to, sera, plasma, urine, saliva, mucous, pleuralfluid, synovial fluid and spinal fluid.

Methods of Ameliorating or Reducing Symptoms of or Treating, orPreventing, Diseases and Disorders Associated with, IL-6

In an embodiment of the invention, IL-6 antagonists described herein areuseful for ameliorating or reducing the symptoms of, or treating, orpreventing, diseases and disorders associated with IL-6. IL-6antagonists described herein can also be administered in atherapeutically effective amount to patients in need of treatment ofdiseases and disorders associated with IL-6 in the form of apharmaceutical composition as described in greater detail below.

In one embodiment of the invention, IL-6 antagonists described hereinare useful for ameliorating or reducing the symptoms of, or treating, orpreventing, diseases and disorders associated with elevated C-reactiveprotein (CRP). Such diseases include any disease that exhibits chronicinflammation, e.g., rheumatoid arthritis, juvenile rheumatoid arthritis,psoriasis, psoriatic arthropathy, ankylosing spondylitis, systemic lupuserythematosis, Crohn's disease, ulcerative colitis, pemphigus,dermatomyositis, polymyositis, polymyalgia rheumatica, giant cellarteritis, vasculitis, polyarteritis nodosa, Wegener's granulomatosis,Kawasaki disease, isolated CNS vasculitis, Churg-Strauss arteritis,microscopic polyarteritis, microscopic polyangiitis, Henoch-Schonleinpurpura, essential cryoglobulinemic vasculitis, rheumatoid vasculitis,cryoglobulinemia, relapsing polychondritis, Behcet's disease, Takayasu'sarteritis, ischemic heart disease, stroke, multiple sclerosis, sepsis,vasculitis secondary to viral infection (e.g., hepatitis B, hepatitis C,HIV, cytomegalovirus, Epstein-Barr virus, Parvo B19 virus, etc.),Buerger's Disease, cancer, advanced cancer, Osteoarthritis, systemicsclerosis, CREST syndrome, Reiter's disease, Paget's disease of bone,Sjogran's syndrome, diabetes type 1, diabetes type 2, familialMediterrean fever, autoimmune thrombocytopenia, autoimmune hemolyticanemia, autoimmune thyroid diseases, pernicious anemia, vitiligo,alopecia areata, primary biliary cirrhosis, autoimmune chronic activehepatitis, alcoholic cirrhosis, viral hepatitis including hepatitis Band C, other organ specific autoimmune diseases, burns, idiopathicpulmonary fibrosis, chronic obsructive pulmonary disease, allergicasthma, other allergic conditions or any combination thereof.

In one embodiment of the invention, IL-6 antagonists described hereinare useful for ameliorating or reducing the symptoms of, or treating, orpreventing, diseases and disorders associated with reduced serumalbumin, e.g. rheumatoid arthritis, cancer, advanced cancer, liverdisease, renal disease, inflammatory bowel disease, celiac's disease,trauma, burns, other diseases associated with reduced serum albumin, orany combination thereof.

In another embodiment of the invention, IL-6 antagonists describedherein are administered to a patient in combination with another activeagent. For example, IL-6 antagonist may be co-administered with one ormore chemotherapy agents, such as VEGF antagonists, EGFR antagonists,platins, taxols, irinotecan, 5-fluorouracil, gemcytabine, leucovorine,steroids, cyclophosphamide, melphalan, vinca alkaloids (e.g.,vinblastine, vincristine, vindesine and vinorelbine), mustines, tyrosinekinase inhibitors, radiotherapy, sex hormone antagonists, selectiveandrogen receptor modulators, selective estrogen receptor modulators,PDGF antagonists, TNF antagonists, IL-I antagonists, interleukins (e.g.IL-I2 or IL-2), IL-I2R antagonists, Toxin conjugated monoclonalantibodies, tumor antigen specific monoclonal antibodies, Erbitux,Avastin, Pertuzumab, anti-CD20 antibodies, Rituxan, ocrelizumab,ofatumumab, DXL625, herceptin, or any combination thereof.

In one embodiment of the invention, anti-IL-6 antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, diseases and disordersassociated with fatigue. Diseases and disorders associated with fatigueinclude, but are not limited to, general fatigue, exercise-inducedfatigue, cancer-related fatigue, fibromyalgia, inflammatorydisease-related fatigue and chronic fatigue syndrome. See, for example,Esper D H, et al, The cancer cachexia syndrome: a review of metabolicand clinical manifestations, Nutr Clin Pract., 2005 August; 20(4):369-76; Vgontzas A N, et al, IL-6 and its circadian secretion inhumans, Neuroimmunomodulation, 2005; I12(3):131-40; Robson-Ansley, P J,et al, Acute interleukin-6 administration impairs athletic performancein healthy, trained male runners, Can J Appl Physiol., 2004 August;29(4):4II-8; Shephard R J., Cytokine responses to physical activity,with particular reference to IL-6: sources, actions, and clinicalimplications, Crit Rev Immunol., 2002; 22(3):165-82; Arnold, M C, et al,Using an interleukin-6 challenge to evaluate neuropsychologicalperformance in chronic fatigue syndrome, Psychol Med., 2002 August;32(6):1075-89; Kurzrock R., The role of cytokines in cancer-relatedfatigue, Cancer, 2001 Sep. 15; 92(6 Suppl):1684-8; Nishimoto N, et al,Improvement in Castleman's disease by humanized anti-interleukin-6receptor antibody therapy, Blood, 2000 Jan. 1; 95 (1):56-61; Vgontzas AN, et al, Circadian interleukin-6 secretion and quantity and depth ofsleep, J Clin Endocrinol Metab., 1999 August; 84(8):2603-7; andSpath-Schwalbe E, et al, Acute effects of recombinant human interleukin6 on endocrine and central nervous sleep functions in healthy men, JClin Endocrinol Metab., 1998 May; 83(5):1573-9; the disclosures of eachof which are herein incorporated by reference in their entireties.

In a preferred embodiment of the invention, anti-IL-6 antibodiesdescribed herein, or fragments thereof, are useful for ameliorating orreducing the symptoms of, or treating, or preventing, cachexia. Diseasesand disorders associated with cachexia include, but are not limited to,cancer-related cachexia, cardiac-related cachexia, respiratory-relatedcachexia, renal-related cachexia and age-related cachexia. See, forexample, Barton, B E., Interleukin-6 and new strategies for thetreatment of cancer, hyperproliferative diseases and paraneoplasticsyndromes, Expert Opin Ther Targets, 2005 August; 9(4):737-52; Zaki M H,et al, CNTO 328, a monoclonal antibody to IL-6, inhibits humantumor-induced cachexia in nude mice, Int J Cancer, 2004 Sep. 10;111(4):592-5; Trikha M, et al, Targeted anti-interleukin-6 monoclonalantibody therapy for cancer: a review of the rationale and clinicalevidence, Clin Cancer Res., 2003 Oct. 15; 9(13):4653-65; Lelli G, et al,Treatment of the cancer anorexia-cachexia syndrome: a criticalreappraisal, J Chemother., 2003 June; 15(3):220-5; Argiles J M, et al,Cytokines in the pathogenesis of cancer cachexia, Curr Opin Clin NutrMetab Care, 2003 July; 6(4):401-6; Barton B E., IL-6-like cytokines andcancer cachexia: consequences of chronic inflammation, Immunol Res.,2001; 23(1):41-58; Yamashita J I, et al, Medroxyprogesterone acetate andcancer cachexia: interleukin-6 involvement, Breast Cancer, 2000;7(2):130-5; Yeh S S, et al, Geriatric cachexia: the role of cytokines,Am J Clin Nutr., 1999 August; 70(2):183-97; Strassmann G, et al,Inhibition of experimental cancer cachexia by anti-cytokine andanti-cytokine-receptor therapy, Cytokines Mol Ther., 1995 June;1(2):107-13; Fujita J, et al, Anti-interleukin-6 receptor antibodyprevents muscle atrophy in colon-26 adenocarcinoma-bearing mice withmodulation of lysosomal and ATP-ubiquitin-dependent proteolyticpathways, Int J Cancer, 1996 Nov. 27; 68(5):637-43; Tsujinaka T, et al,Interleukin 6 receptor antibody inhibits muscle atrophy and modulatesproteolytic systems in interleukin 6 transgenic mice, J Clin Invest.,1996 Jan. 1; 97(1):244-9; Emilie D, et al, Administration of ananti-interleukin-6 monoclonal antibody to patients with acquiredimmunodeficiency syndrome and lymphoma: effect on lymphoma growth and onB clinical Symptoms, Blood, 1994 Oct. 15; 84 (8):2472-9; and StrassmannG, et al, Evidence for the involvement of interleukin 6 in experimentalcancer cachexia, J Clin Invest., 1992 May; 89(5):1681-4; the disclosuresof each of which are herein incorporated by reference in theirentireties.

In another embodiment of the invention, anti-IL-6 antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, autoimmune diseases anddisorders. Diseases and disorders associated with autoimmunity include,but are not limited to, rheumatoid arthritis, systemic lupuserythematosis (SLE), systemic juvenile idiopathic arthritis, psoriasis,psoriatic arthropathy, ankylosing spondylitis, inflammatory boweldisease (IBD), polymyalgia rheumatica, giant cell arteritis, autoimmunevasculitis, graft versus host disease (GVHD), Sjogren's syndrome, adultonset Still's disease. In a preferred embodiment of the invention,humanized anti-IL-6 antibodies described herein, or fragments thereof,are useful for ameliorating or reducing the symptoms of, or treating, orpreventing, rheumatoid arthritis and systemic juvenile idiopathicarthritis. See, for example, Nishimoto N., Clinical studies in patientswith Castleman's disease, Crohn's disease, and rheumatoid arthritis inJapan, Clin Rev Allergy Immunol., 2005 June; 28(3):221-30; Nishimoto N,et al, Treatment of rheumatoid arthritis with humanizedanti-interleukin-6 receptor antibody: a multicenter, double-blind,placebo-controlled trial, Arthritis Rheum., 2004 June; 50(6):1761-9;Choy E., Interleukin 6 receptor as a target for the treatment ofrheumatoid arthritis, Ann Rheum Dis., 2003 November; 62 Suppl 2:ii68-9;Nishimoto N, et al, Toxicity, pharmacokinetics, and dose-finding studyof repetitive treatment with the humanized anti-interleukin 6 receptorantibody MRA in rheumatoid arthritis. Phase I/II clinical study, JRheumatol., 2003 July; 30(7):1426-35; Mihara M, et al, Humanizedantibody to human interleukin-6 receptor inhibits the development ofcollagen arthritis in cynomolgus monkeys, Clin Immunol., 2001 March;98(3):319-26; Nishimoto N, et al, Anti-interleukin 6 receptor antibodytreatment in rheumatic disease, Ann Rheum Dis., 2000 November; 59 Suppl1:i21-7; Tackey E, et al, Rationale for interleukin-6 blockade insystemic lupus erythematosus, Lupus, 2004; 13(5):339-43; Finck B K, etal, Interleukin 6 promotes murine lupus in NZB/NZW F1 mice, J ClinInvest., 1994 August; 94 (2):585-91; Kitani A, et al, Autostimulatoryeffects of IL-6 on excessive B cell differentiation in patients withsystemic lupus erythematosus: analysis of IL-6 production and IL-6Rexpression, Clin Exp Immunol., 1992 April; 88(1):75-83; Stuart R A, etal, Elevated serum interleukin-6 levels associated with active diseasein systemic connective tissue disorders, Clin Exp Rheumatol., 1995January-February; 13 (1):17-22; Mihara M, et al, IL-6 receptor blockageinhibits the onset of autoimmune kidney disease in NZB/W F1 mice, ClinExp Immunol., 1998 June; 12(3):397-402; Woo P, et al, Open label phaseII trial of single, ascending doses of MRA in Caucasian children withsevere systemic juvenile idiopathic arthritis: proof of principle of theefficacy of IL-6 receptor blockade in this type of arthritis anddemonstration of prolonged clinical improvement, Arthritis Res Ther.,2005; 7(6):RI281-8. Epub 2005 Sep. 15; Yokota S, et al, Clinical studyof tocilizumab in children with systemic-onset juvenile idiopathicarthritis, Clin Rev Allergy Immunol., 2005 June; 28(3):231-8; Yokota S,et al, Therapeutic efficacy of humanized recombinant anti-interleukin-6receptor antibody in children with systemic-onset juvenile idiopathicarthritis, Arthritis Rheum., 2005 March; 52(3):818-25; de Benedetti F,et al, Targeting the interleukin-6 receptor: a new treatment forsystemic juvenile idiopathic arthritis?, Arthritis Rheum., 2005 March;52(3):687-93; De Benedetti F, et al, Is systemic juvenile rheumatoidarthritis an interleukin 6 mediated disease?, J Rheumatol., 1998February; 25(2):203-7; Ishihara K, et al, IL-6 in autoimmune disease andchronic inflammatory proliferative disease, Cytokine Growth Factor Rev.,2002 August-October; 13 (4-5):357-68; Gilhar A, et al, In vivo effectsof cytokines on psoriatic skin grafted on nude mice: involvement of thetumor necrosis factor (TNF) receptor, Clin Exp Immunol., 1996 October;106(1):134-42; Spadaro A, et al, Interleukin-6 and soluble interleukin-2receptor in psoriatic arthritis: correlations with clinical andlaboratory parameters, Clin Exp Rheumatol., 1996 July-August; 14(4):413-6; Ameglio F, et al, Interleukin-6 and tumor necrosis factorlevels decrease in the suction blister fluids of psoriatic patientsduring effective therapy, Dermatology, 1994; 189(4):359-63; Wendling D,et al, Combination therapy of anti-CD4 and anti-IL-6 monoclonalantibodies in a case of severe spondylarthropathy, Br J Rheumatol., 1996December; 35(12):1330; Gratacos J, et al, Serum cytokines (IL-6,TNF-alpha, IL-1 beta and IFN-gamma) in ankylosing spondylitis: a closecorrelation between serum IL-6 and disease activity and severity, Br JRheumatol., 1994 October; 33(10):927-31; Ito H., Treatment of Crohn'sdisease with anti-IL-6 receptor antibody, J Gastroenterol., 2005 March;40 Suppl 16:32-4; Ito H, et al, A pilot randomized trial of a humananti-interleukin-6 receptor monoclonal antibody in active Crohn'sdisease, Gastroenterology, 2004 April; 126(4):989-96; discussion 947;Ito H., IL-6 and Crohn's disease, Curr Drug Targets Inflamm Allergy,2003 June; 2(2):12530; Ito H, et al, Anti-IL-6 receptor monoclonalantibody inhibits leukocyte recruitment and promotes T-cell apoptosis ina murine model of Crohn's disease, J Gastroenterol., 2002 November; 37Suppl 14:56-61; Ito H., Anti-interleukin-6 therapy for Crohn's disease,Curr Pharm Des., 2003; 9(4):295-305; Salvarani C, et al, Acute-phasereactants and the risk of relapse/recurrence in polymyalgia rheumatica:a prospective follow-up study, Arthritis Rheum., 2005 Feb. 15;53(1):33-8; Roche N E, et al, Correlation of interleukin-6 productionand disease activity in polymyalgia rheumatica and giant cell arteritis,Arthritis Rheum., 1993 September; 36(9):1286-94; Gupta M, et al,Cytokine modulation with immune gamma-globulin in peripheral blood ofnormal children and its implications in Kawasaki disease treatment, JClin Immunol., 2001 May; 21(3):193-9; Noris M, et al, Interleukin-6 andRANTES in Takayasu arteritis: a guide for therapeutic decisions?,Circulation, 1999 Jul. 6; 100(1):55-60; Besbas N, et al, The role ofcytokines in Henoch Schonlein purpura, Scand J Rheumatol., 1997;26(6):456-60; Hirohata S, et al, Cerebrospinal fluid interleukin-6 inprogressive Neuro-Behcet's syndrome, Clin Immunol Immunopathol., 1997January; 82(1):12-7; Yamakawa Y, et al, Interleukin-6 (IL-6) in patientswith Behcet's disease, J Dermatol Sci., 1996 March; 11(3):189-95; Kim DS., Serum interleukin-6 in Kawasaki disease, Yonsei Med J., 1992 June;33(2):183-8; Lange, A., et al, Cytokines, adhesion molecules (E-selectinand VCAM-1) and graft-versus-host disease, Arch. Immunol Ther Exp.,1995, 43(2):99-105; Tanaka, J., et al, Cytokine gene expression afterallogeneic bone marrow transplantation, Leuk. Lymphoma, 199516(5-6):413-418; Dickenson, A M, et al, Predicting outcome inhematological stem cell transplantation, Arch Immunol Ther Exp., 200250(6):371-8; Zeiser, R, et al, Immunopathogenesis of acutegraft-versus-host disease: implications for novel preventive andtherapeutic strategies, Ann Hematol., 2004 83(9):551-65; Dickinson, A M,et al, Genetic polymorphisms predicting the outcome of bone marrowtransplants, Br. J Haematol., 2004 127(5):479-90; and Scheinberg M A, etal, Interleukin 6: a possible marker of disease activity in adult onsetStill's disease, Clin Exp Rheumatol., 1996 November-December; 14(6):653-5, the disclosures of each of which are herein incorporated byreference in their entireties.

In another embodiment of the invention, anti-IL-6 antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, diseases and disordersassociated with the skeletal system. Diseases and disorders associatedwith the skeletal system include, but are not limited to,osteoarthritis, osteoporosis and Paget's disease of bone. In a preferredembodiment of the invention, humanized anti-IL-6 antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, osteoarthritis. See, forexample, Malemud C J., Cytokines as therapeutic targets forosteoarthritis, BioDrugs, 2004; 18(1):23-35; Westacott C I, et al,Cytokines in osteoarthritis: mediators or markers of joint destruction?,Semin Arthritis Rheum., 1996 February; 25(4):254-72; Sugiyama T.,Involvement of interleukin-6 and prostaglandin E2 in particularosteoporosis of postmenopausal women with rheumatoid arthritis, J BoneMiner Metab., 2001; 19(2):89-96; Abrahamsen B, et al, Cytokines and boneloss in a 5-year longitudinal study-hormone replacement therapysuppresses serum soluble interleukin-6 receptor and increasesinterleukin-1-receptor antagonist: the Danish Osteoporosis PreventionStudy, J Bone Miner Res., 2000 August; 15(8):1545-54; Straub R H, et al,Hormone replacement therapy and interrelation between seruminterleukin-6 and body mass index in postmenopausal women: apopulation-based study, J Clin Endocrinol Metab., 2000 March;85(3):1340-4; Manolagas S C, The role of IL-6 type cytokines and theirreceptors in bone, Ann N Y Acad Sci., 1998 May 1; 840:194-204; Ershler WB, et al, Immunologic aspects of osteoporosis, Dev Comp Immunol., 1997November-December; 21(6):487-99; Jilka R L, et al, Increased osteoclastdevelopment after estrogen loss: mediation by interleukin-6, Science,1992 Jul. 3; 257(5066):88-91; Kallen K J, et al, New developments inIL-6 dependent biology and therapy: where do we stand and what are theoptions?, Expert Opin Investig Drugs, 1999 September; 8(9):1327-49;Neale S D, et al, The influence of serum cytokines and growth factors onosteoclast formation in Paget's disease, Q J M, 2002 April; 95(4):233-40; Roodman G D, Osteoclast function In Paget's disease andmultiple myeloma, Bone, 1995 August; 17(2 Suppl):57S-61S; Hoyland J A,et al, Interleukin-6, IL-6 receptor, and IL-6 nuclear factor geneexpression in Paget's disease, J Bone Miner Res., 1994 January;9(1):75-80; and Roodman G D, et al, Interleukin 6. A potentialautocrine/paracrine factor in Paget's disease of bone, J Clin Invest.,1992 January; 89(1):46-52; the disclosures of each of which are hereinincorporated by reference in their entireties.

In another embodiment of the invention, anti-IL-6 antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, diseases and disordersassociated with cancer. Diseases and disorders associated with cancerinclude, but are not limited to, Acanthoma, Acinic cell carcinoma,Acoustic neuroma, Acral lentiginous melanoma, Acrospiroma, Acuteeosinophilic leukemia, Acute lymphoblastic leukemia, Acutemegakaryoblastic leukemia, Acute monocytic leukemia, Acute myeloblasticleukemia with maturation, Acute myeloid dendritic cell leukemia, Acutemyeloid leukemia, Acute promyelocytic leukemia, Adamantinoma,Adenocarcinoma, Adenoid cystic carcinoma, Adenoma, Adenomatoidodontogenic tumor, Adrenocortical carcinoma, Adult T-cell leukemia,Aggressive NK-cell leukemia, AIDS-Related Cancers, AIDS-relatedlymphoma, Alveolar soft part sarcoma, Ameloblastic fibroma, Anal cancer,Anaplastic large cell lymphoma, Anaplastic thyroid cancer,Angioimmunoblastic T-cell lymphoma, Angiomyolipoma, Angiosarcoma,Appendix cancer, Astrocytoma, Atypical teratoid rhabdoid tumor, Basalcell carcinoma, Basal-like carcinoma, B-cell leukemia, B-cell lymphoma,Bellini duct carcinoma, Biliary tract cancer, Bladder cancer, Blastoma,Bone Cancer, Bone tumor, Brain Stem Glioma, Brain Tumor, Breast Cancer,Brenner tumor, Bronchial Tumor, Bronchioloalveolar carcinoma, Browntumor, Burkitt's lymphoma, Cancer of Unknown Primary Site, CarcinoidTumor, Carcinoma, Carcinoma in situ, Carcinoma of the penis, Carcinomaof Unknown Primary Site, Carcinosarcoma, Castleman's Disease, CentralNervous System Embryonal Tumor, Cerebellar Astrocytoma, CerebralAstrocytoma, Cervical Cancer, Cholangiocarcinoma, Chondroma,Chondrosarcoma, Chordoma, Choriocarcinoma, Choroid plexus papilloma,Chronic Lymphocytic Leukemia, Chronic monocytic leukemia, Chronicmyelogenous leukemia, Chronic Myeloproliferative Disorder, Chronicneutrophilic leukemia, Clear-cell tumor, Colon Cancer, Colorectalcancer, Craniopharyngioma, Cutaneous T-cell lymphoma, Degos disease,Dermatofibrosarcoma protuberans, Dermoid cyst, Desmoplastic small roundcell tumor, Diffuse large B cell lymphoma, Dysembryoplasticneuroepithelial tumor, Embryonal carcinoma, Endodermal sinus tumor,Endometrial cancer, Endometrial Uterine Cancer, Endometrioid tumor,Enteropathy-associated T-cell lymphoma, Ependymoblastoma, Ependymoma,Epithelioid sarcoma, Erythroleukemia, Esophageal cancer,Esthesioneuroblastoma, Ewing Family of Tumor, Ewing Family Sarcoma,Ewing's sarcoma, Extracranial Germ Cell Tumor, Extragonadal Germ CellTumor, Extrahepatic Bile Duct Cancer, Extramammary Paget's disease,Fallopian tube cancer, Fetus in fetu, Fibroma, Fibrosarcoma, Follicularlymphoma, Follicular thyroid cancer, Gallbladder Cancer, Gallbladdercancer, Ganglioglioma, Ganglioneuroma, Gastric Cancer, Gastric lymphoma,Gastrointestinal cancer, Gastrointestinal Carcinoid Tumor,Gastrointestinal Stromal Tumor, Gastrointestinal stromal tumor, Germcell tumor, Germinoma, Gestational choriocarcinoma, GestationalTrophoblastic Tumor, Giant cell tumor of bone, Glioblastoma multiforme,Glioma, Gliomatosis cerebri, Glomus tumor, Glucagonoma, Gonadoblastoma,Granulosa cell tumor, Hairy Cell Leukemia, Hairy cell leukemia, Head andNeck Cancer, Head and neck cancer, Heart cancer, Hemangioblastoma,Hemangiopericytoma, Hemangiosarcoma, Hematological malignancy,Hepatocellular carcinoma, Hepatosplenic T-cell lymphoma, Hereditarybreast-ovarian cancer syndrome, Hodgkin Lymphoma, Hodgkin's lymphoma,Hypopharyngeal Cancer, Hypothalamic Glioma, Inflammatory breast cancer,Intraocular Melanoma, Islet cell carcinoma, Islet Cell Tumor, Juvenilemyelomonocytic leukemia, Kaposi Sarcoma, Kaposi's sarcoma, KidneyCancer, Klatskin tumor, Krukenberg tumor, Laryngeal Cancer, Laryngealcancer, Lentigo maligna melanoma, Leukemia, Leukemia, Lip and OralCavity Cancer, Liposarcoma, Lung cancer, Luteoma, Lymphangioma,Lymphangiosarcoma, Lymphoepithelioma, Lymphoid leukemia, Lymphoma,Macroglobulinemia, Malignant Fibrous Histiocytoma, Malignant fibroushistiocytoma, Malignant Fibrous Histiocytoma of Bone, Malignant Glioma,Malignant Mesothelioma, Malignant peripheral nerve sheath tumor,Malignant rhabdoid tumor, Malignant triton tumor, MALT lymphoma, Mantlecell lymphoma, Mast cell leukemia, Mediastinal germ cell tumor,Mediastinal tumor, Medullary thyroid cancer, Medulloblastoma,Medulloblastoma, Medulloepithelioma, Melanoma, Melanoma, Meningioma,Merkel Cell Carcinoma, Mesothelioma, Mesothelioma, Metastatic SquamousNeck Cancer with Occult Primary, Metastatic urothelial carcinoma, MixedMüllerian tumor, Monocytic leukemia, Mouth Cancer, Mucinous tumor,Multiple Endocrine Neoplasia Syndrome, Multiple Myeloma, Multiplemyeloma, Mycosis Fungoides, Mycosis fungoides, Myelodysplastic Disease,Myelodysplastic Syndromes, Myeloid leukemia, Myeloid sarcoma,Myeloproliferative Disease, Myxoma, Nasal Cavity Cancer, NasopharyngealCancer, Nasopharyngeal carcinoma, Neoplasm, Neurinoma, Neuroblastoma,Neuroblastoma, Neurofibroma, Neuroma, Nodular melanoma, Non-HodgkinLymphoma, Non-Hodgkin lymphoma, Nonmelanoma Skin Cancer, Non-Small CellLung Cancer, Ocular oncology, Oligoastrocytoma, Oligodendroglioma,Oncocytoma, Optic nerve sheath meningioma, Oral Cancer, Oral cancer,Oropharyngeal Cancer, Osteosarcoma, Osteosarcoma, Ovarian Cancer,Ovarian cancer, Ovarian Epithelial Cancer, Ovarian Germ Cell Tumor,Ovarian Low Malignant Potential Tumor, Paget's disease of the breast,Pancoast tumor, Pancreatic Cancer, Pancreatic cancer, Papillary thyroidcancer, Papillomatosis, Paraganglioma, Paranasal Sinus Cancer,Parathyroid Cancer, Penile Cancer, Perivascular epithelioid cell tumor,Pharyngeal Cancer, Pheochromocytoma, Pineal Parenchymal Tumor ofIntermediate Differentiation, Pineoblastoma, Pituicytoma, Pituitaryadenoma, Pituitary tumor, Plasma Cell Neoplasm, Pleuropulmonaryblastoma, Polyembryoma, Precursor T-lymphoblastic lymphoma, Primarycentral nervous system lymphoma, Primary effusion lymphoma, PrimaryHepatocellular Cancer, Primary Liver Cancer, Primary peritoneal cancer,Primitive neuroectodermal tumor, Prostate cancer, Pseudomyxomaperitonei, Rectal Cancer, Renal cell carcinoma, Respiratory TractCarcinoma Involving the NUT Gene on Chromosome 15, Retinoblastoma,Rhabdomyoma, Rhabdomyosarcoma, Richter's transformation, Sacrococcygealteratoma, Salivary Gland Cancer, Sarcoma, Schwannomatosis, Sebaceousgland carcinoma, Secondary neoplasm, Seminoma, Serous tumor,Sertoli-Leydig cell tumor, Sex cord-stromal tumor, Sezary Syndrome,Signet ring cell carcinoma, Skin Cancer, Small blue round cell tumor,Small cell carcinoma, Small Cell Lung Cancer, Small cell lymphoma, Smallintestine cancer, Soft tissue sarcoma, Somatostatinoma, Soot wart,Spinal Cord Tumor, Spinal tumor, Splenic marginal zone lymphoma,Squamous cell carcinoma, Stomach cancer, Superficial spreading melanoma,Supratentorial Primitive Neuroectodermal Tumor, Surfaceepithelial-stromal tumor, Synovial sarcoma, T-cell acute lymphoblasticleukemia, T-cell large granular lymphocyte leukemia, T-cell leukemia,T-cell lymphoma, T-cell prolymphocytic leukemia, Teratoma, Terminallymphatic cancer, Testicular cancer, Thecoma, Throat Cancer, ThymicCarcinoma, Thymoma, Thyroid cancer, Transitional Cell Cancer of RenalPelvis and Ureter, Transitional cell carcinoma, Urachal cancer, Urethralcancer, Urogenital neoplasm, Uterine sarcoma, Uveal melanoma, VaginalCancer, Verner Morrison syndrome, Verrucous carcinoma, Visual PathwayGlioma, Vulvar Cancer, Waldenström's macroglobulinemia, Warthin's tumor,Wilms' tumor, or any combination thereof, as well as drug resistance incancer chemotherapy and cancer chemotherapy toxicity. See, for example,Hirata T, et al, Humanized anti-interleukin-6 receptor monoclonalantibody induced apoptosis of fresh and cloned human myeloma cells invitro, Leuk Res., 2003 April; 27(4):343-9, Bataille R, et al, Biologiceffects of anti-interleukin-6 murine monoclonal antibody in advancedmultiple myeloma, Blood, 1995 Jul. 15; 86 (2):685-91; Goto H, et al,Mouse anti-human interleukin-6 receptor monoclonal antibody inhibitsproliferation of fresh human myeloma cells in vitro, Jpn J Cancer Res.,1994 September; 85(9):958-65; Klein B, et al, Murine anti-interleukin-6monoclonal antibody therapy for a patient with plasma cell leukemia,Blood, 1991 Sep. 1; 78(5):1198-204; Mauray S, et al, Epstein-Barrvirus-dependent lymphoproliferative disease: critical role of IL-6, EurJ Immunol., 2000 July; 30(7):2065-73; Tsunenari T, et al, New xenograftmodel of multiple myeloma and efficacy of a humanized antibody againsthuman interleukin-6 receptor, Blood, 1997 Sep. 15; 90(6):2437-44; EmilieD, et al, Interleukin-6 production in high-grade B lymphomas:correlation with the presence of malignant immunoblasts in acquiredimmunodeficiency syndrome and in human immunodeficiencyvirus-seronegative patients, Blood, 1992 Jul. 15; 80(2):498-504; EmilieD, et al, Administration of an anti-interleukin-6 monoclonal antibody topatients with acquired immunodeficiency syndrome and lymphoma: effect onlymphoma growth and on B clinical Symptoms, Blood, 1994 Oct. 15;84(8):2472-9; Smith P C, et al, Anti-interleukin-6 monoclonal antibodyinduces regression of human prostate cancer xenografts in nude mice,Prostate, 2001 Jun. 15; 48(1):47-53; Smith P C, et al, Interleukin-6 andprostate cancer progression, Cytokine Growth Factor Rev., 2001 March;12(1):33-40; Chung T D, et al, Characterization of the role of IL-6 inthe progression of prostate cancer, Prostate, 1999 Feb. 15;38(3):199-207; Okamoto M, et al, Interleukin-6 as a paracrine andautocrine growth factor in human prostatic carcinoma cells in vitro,Cancer Res., 1997 January 1; 57(1):141-6; Reittie J E, et al,Interleukin-6 inhibits apoptosis and tumor necrosis factor inducedproliferation of B-chronic lymphocytic leukemia, Leuk Lymphoma, 1996June; 22(1-2):83-90, follow 186, color plate VI; Sugiyama H, et al, Theexpression of IL-6 and its related genes in acute leukemia, LeukLymphoma, 1996 March; 21(1-2):49-52; Bataille R, et al, Effects of ananti-interleukin-6 (IL-6) murine monoclonal antibody in a patient withacute monoblastic leukemia, Med Oncol Tumor Pharmacother., 1993;10(4):185-8; Kedar I, et al, Thalidomide reduces serum C-reactiveprotein and interleukin-6 and induces response to IL-2 in a fraction ofmetastatic renal cell cancer patients who failed IL-2-based therapy, IntJ Cancer, 2004 Jun. 10; 110(2):260-5; Angelo L S, Talpaz M, Kurzrock R,Autocrine interleukin-6 production in renal cell carcinoma: evidence forthe involvement of p53, Cancer Res., 2002 Feb. 1; 62(3):932-40;Nishimoto N, Humanized anti-interleukin-6 receptor antibody treatment ofmulticentric Castleman disease, Blood, 2005 Oct. 15; 106(8):2627-32,Epub 2005 Jul. 5; Katsume A, et al, Anti-interleukin 6 (IL-6) receptorantibody suppresses Castleman's disease like symptoms emerged in IL-6transgenic mice, Cytokine, 2002 Dec. 21; 20(6):304-11; Nishimoto N, etal, Improvement in Castleman's disease by humanized anti-interleukin-6receptor antibody therapy, Blood, 2000 January 1; 95(1):56-61; ScrepantiI, Inactivation of the IL-6 gene prevents development of multicentricCastleman's disease in C/EBP beta-deficient mice, J Exp Med., 1996 Oct.1; 184(4):1561-6; Hsu S M, et al, Expression of interleukin-6 inCastleman's disease, Hum Pathol., 1993 August; 24(8):833-9; Yoshizaki K,et al, Pathogenic significance of interleukin-6 (IL 6/BSF-2) inCastleman's disease, Blood, 1989 September; 74(4):1360-7; Nilsson M B,et al, Interleukin-6, secreted by human ovarian carcinoma cells, is apotent proangiogenic cytokine, Cancer Res., 2005 Dec. 1;65(23):10794-800; Toutirais O, et al, Constitutive expression ofTGF-betal, interleukin-6 and interleukin-8 by tumor cells as a majorcomponent of immune escape in human ovarian carcinoma, Eur CytokineNetw., 2003 October-December; 14(4):246-55; Obata N H, et al, Effects ofinterleukin 6 on in vitro cell attachment, migration and invasion ofhuman ovarian carcinoma, Anticancer Res., 1997 January-February; 17(1A):337-42; Dedoussis G V, et al, Endogenous interleukin 6 conveysresistance to cis-diamminedichloroplatinum-mediated apoptosis of theK562 human leukemic cell line, Exp Cell Res., 1999 Jun. 15;249(2):269-78; Borsellino N, et al, Blocking signaling through the Gp130receptor chain by interleukin-6 and oncostatin M inhibits PC-3 cellgrowth and sensitizes the tumor cells to etoposide andcisplatin-mediated cytotoxicity, Cancer, 1999 Jan. 1; 85(1):134-44;Borsellino N, et al, Endogenous interleukin 6 is a resistance factor forcis-diamminedichloroplatinum and etoposide-mediated cytotoxicity ofhuman prostate carcinoma cell lines, Cancer Res., 1995 Oct. 15;55(20):4633-9; Mizutani Y, et al, Sensitization of human renal cellcarcinoma cells to cis-diamminedichloroplatinum(II) by anti-interleukin6 monoclonal antibody or anti-interleukin 6 receptor monoclonalantibody; Cancer Res., 1995 Feb. 1; 55(3):590-6; Yusuf R Z, et al,Paclitaxel resistance: molecular mechanisms and pharmacologicmanipulation, Curr Cancer Drug Targets, 2003 February; 3(1):1-19; DuanZ, et al, Overexpression of IL-6 but not IL-8 increases paclitaxelresistance of U-20S human osteosarcoma cells, Cytokine, 2002 Mar. 7;17(5):234-42; Conze D, et al, Autocrine production of interleukin 6causes multidrug resistance in breast cancer cells, Cancer Res., 2001Dec. 15; 61(24):8851-8; Rossi J F, et al, Optimizing the use ofanti-interleukin-6 monoclonal antibody with dexamethasone and 140 mg/m2of melphalan in multiple myeloma: results of a pilot study includingbiological aspects, Bone Marrow Transplant, 2005 November; 36(9):771-9;and Tonini G, et al, Oxaliplatin may induce cytokine-release syndrome incolorectal cancer patients, J Biol Regul Homeost Agents, 2002April-June; 16 (2):105-9; the disclosures of each of which are hereinincorporated by reference in their entireties.

In another embodiment of the invention, anti-IL-6 antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, ischemic heart disease,atherosclerosis, obesity, diabetes, asthma, multiple sclerosis,Alzheimer's disease, cerebrovascular disease, fever, acute phaseresponse, allergies, anemia, anemia of inflammation (anemia of chronicdisease), hypertension, depression, depression associated with a chronicillness, thrombosis, thrombocytosis, acute heart failure, metabolicsyndrome, miscarriage, obesity, chronic prostatitis, glomerulonephritis,pelvic inflammatory disease, reperfusion injury, and transplantrejection. See, for example, Tzoulaki I, et al, C-reactive protein,interleukin-6, and soluble adhesion molecules as predictors ofprogressive peripheral atherosclerosis in the general population:Edinburgh Artery Study, Circulation, 2005 Aug. 16; 112(7):976-83, Epub2005 Aug. 8; Rattazzi M, et al, C-reactive protein and interleukin-6 invascular disease: culprits or passive bystanders?, J Hypertens., 2003October; 21(10):1787-803; Ito T, et al, HMG-CoA reductase inhibitorsreduce interleukin-6 synthesis in human vascular smooth muscle cells,Cardiovasc Drugs Ther., 2002 March; 16(2):121-6; Stenvinkel P, et al,Mortality, malnutrition, and atherosclerosis in ESRD: what is the roleof interleukin-6?, Kidney Int Suppl., 2002 May; (80):103-8; Yudkin J S,et al, Inflammation, obesity, stress and coronary heart disease: isinterleukin-6 the link?, Atherosclerosis, 2000 February; 148(2):209-14;Huber S A, et al, Interleukin-6 exacerbates early atherosclerosis inmice, Arterioscler Thromb Vasc Biol., 1999 October; 19(10):2364-7; KadoS, et al, Circulating levels of interleukin-6, its soluble receptor andinterleukin-6/interleukin-6 receptor complexes in patients with type 2diabetes mellitus, Acta Diabetol., 1999 June; 36(1-2):67-72; Sukovich DA, et al, Expression of interleukin-6 in atherosclerotic lesions of maleApoE-knockout mice: inhibition by 17beta-estradiol, Arterioscler ThrombVasc Biol., 1998 September; 8(9):1498-505; Klover P J, et al,Interleukin-6 depletion selectively improves hepatic insulin action inobesity, Endocrinology, 2005 August; 146(8):3417-27, Epub 2005 Apr. 21;Lee Y H, et al, The evolving role of inflammation in obesity and themetabolic syndrome, Curr Diab Rep., 2005 February; 5(1):70-5; Diamant M,et al, The association between abdominal visceral fat and carotidstiffness is mediated by circulating inflammatory markers inuncomplicated type 2 diabetes, J Clin Endocrinol Metab., 2005 March;90(3):1495-501, Epub 2004 Dec. 21; Bray G A, Medical consequences ofobesity, J Clin Endocrinol Metab., 2004 June; 89(6):2583 9; Klover P J,et al, Chronic exposure to interleukin-6 causes hepatic insulinresistance in mice, Diabetes, 2003 November; 52 (11):2784-9; Yudkin J S,et al, Inflammation, obesity, stress and coronary heart disease: isinterleukin-6 the link?, Atherosclerosis, 2000 February; 148(2):209-14;Doganci A, et al, Pathological role of IL-6 in the experimental allergicbronchial asthma in mice, Clin Rev Allergy Immunol., 2005 June;28(3):257-70; Doganci A, et al, The IL-6R alpha chain controls lungCD4+CD25+ Treg development and function during allergic airwayinflammation in vivo, J Clin Invest., 2005 February; 115(2):313 25,(Erratum in: J Clin Invest., 2005 May; 115(5):1388, Lehr, Hans A[added]); Stelmasiak Z, et al, IL 6 and sIL-6R concentration in thecerebrospinal fluid and serum of MS patients, Med Sci Monit., 2001September-October; 7(5):914-8; Tilgner J, et al, Continuousinterleukin-6 application in vivo via macroencapsulation ofinterleukin-6-expressing COS-7 cells induces massive gliosis, Glia, 2001September; 35(3):234-45, Brunello A G, et al, Astrocytic alterations ininterleukin-6 Soluble interleukin-6 receptor alpha double-transgenicmice, Am J Pathol., 2000 November; 157(5):1485-93; Hampel H, et al,Pattern of interleukin-6 receptor complex immunoreactivity betweencortical regions of rapid autopsy normal and Alzheimer's disease brain,Eur Arch Psychiatry Clin Neurosci., 2005 August; 255(4):269-78, Epub2004 Nov. 26; Cacquevel M, et al, Cytokines in neuroinflammation andAlzheimer's disease, Curr Drug Targets, 2004 August; 5(6):529-34;Quintanilla R A, et al, Interleukin 6 induces Alzheimer-typephosphorylation of tau protein by deregulating the cdk5/p35 pathway, ExpCell Res., 2004 Apr. 15; 295 (1):245-57; Gadient R A, et al,Interleukin-6 (IL-6)—a molecule with both beneficial and destructivepotentials, Prog Neurobiol., 1997 August; 52(5):379-90; Hull M, et al,Occurrence of interleukin-6 in cortical plaques of Alzheimer's diseasepatients may precede transformation of diffuse into neuritic plaques,Ann N Y Acad Sci., 1996 Jan. 17; 777:205-12; Rallidis L S, et al,Inflammatory markers and in-hospital mortality in acute ischaemicstroke, Atherosclerosis, 2005 Dec. 30; Emsley H C, et al, Interleukin-6and acute ischaemic stroke, Acta Neurol Scand., 2005 October;112(4):273-4; Smith C J, et al, Peak plasma interleukin-6 and otherperipheral markers of inflammation in the first week of ischaemic strokecorrelate with brain infarct volume, stroke severity and long-termoutcome, BMC Neurol., 2004 Jan. 15; 4:2; Vila N, et al, Proinflammatorycytokines and early neurological worsening in ischemic stroke, Stroke,2000 October; 31(10):2325-9; and Tarkowski E, et al, Early intrathecalproduction of interleukin-6 predicts the size of brain lesion in stroke,Stroke, 1995 August; 26(8):1393-8; the disclosures of each of which areherein incorporated by reference in their entireties.

In another embodiment of the invention, anti-IL-6 antibodies describedherein, or fragments thereof, are useful for ameliorating or reducingthe symptoms of, or treating, or preventing, diseases and disordersassociated with cytokine storm. Diseases and disorders associated withcytokine storm include, but are not limited to, graft versus hostdisease (GVHD), avian influenza, smallpox, pandemic influenza, adultrespiratory distress syndrome (ARDS), severe acute respiratory syndrome(SARS), sepsis, and systemic inflammatory response syndrome (SIRS). See,for example, Cecil, R. L., Goldman, L., & Bennett, J. C. (2000). Ceciltextbook of medicine. Philadelphia: W. B. Saunders; Ferrara J L, et al.,Cytokine storm of graft-versus-host disease: a critical effector rolefor interleukin-1, Transplant Proc. 1993 February; 25(1 Pt 2):1216-7;Osterholm M T, Preparing for the Next Pandemic, N Engl J Med. 2005 May5; 352(18):1839-42; Huang K J, et al., An interferon-gamma-relatedcytokine storm in SARS patients, J Med Virol. 2005 February;75(2):185-94; and Cheung C Y, et al., Induction of proinflammatorycytokines in human macrophages by influenza A (H5N1) viruses: amechanism for the unusual severity of human disease? Lancet. 2002 Dec.7; 360(9348):1831-7.

In another embodiment of the invention, anti-IL-6 antibodies describedherein, or fragments thereof, are useful as a wakefulness aid.

Administration

In one embodiment of the invention, the anti-IL-6 antibodies describedherein, or IL-6 binding fragments thereof, as well as combinations ofsaid antibody fragments, are administered to a subject at aconcentration of between about 0.1 and 20 mg/kg, such as about 0.4mg/kg, about 0.8 mg/kg, about 1.6 mg/kg, or about 4 mg/kg, of bodyweight of recipient subject. In a preferred embodiment of the invention,the anti-IL-6 antibodies described herein, or IL-6 binding fragmentsthereof, as well as combinations of said antibody fragments, areadministered to a subject at a concentration of about 0.4 mg/kg of bodyweight of recipient subject. In a preferred embodiment of the invention,the anti-IL-6 antibodies described herein, or IL-6 binding fragmentsthereof, as well as combinations of said antibody fragments, areadministered to a recipient subject with a frequency of once everytwenty-six weeks or less, such as once every sixteen weeks or less, onceevery eight weeks or less, or once every four weeks, or less. In anotherpreferred embodiment of the invention, the anti-IL-6 antibodiesdescribed herein, or IL-6 binding fragments thereof, as well ascombinations thereof, are administered to a recipient subject with afrequency at most once per period of approximately one week, such as atmost once per period of approximately two weeks, such as at most onceper period of approximately four weeks, such as at most once per periodof approximately eight weeks, such as at most once per period ofapproximately twelve weeks, such as at most once per period ofapproximately sixteen weeks, such as at most once per period ofapproximately twenty-four weeks.

It is understood that the effective dosage may depend on recipientsubject attributes, such as, for example, age, gender, pregnancy status,body mass index, lean body mass, condition or conditions for which thecomposition is given, other health conditions of the recipient subjectthat may affect metabolism or tolerance of the composition, levels ofIL-6 in the recipient subject, and resistance to the composition (forexample, arising from the patient developing antibodies against thecomposition). A person of skill in the art would be able to determine aneffective dosage and frequency of administration through routineexperimentation, for example guided by the disclosure herein and theteachings in Goodman, L. S., Gilman, A., Brunton, L. L., Lazo, J. S., &Parker, K. L. (2006). Goodman & Gilman's the pharmacological basis oftherapeutics. New York: McGraw-Hill; Howland, R. D., Mycek, M. J.,Harvey, R. A., Champe, P. C., & Mycek, M. J. (2006). Pharmacology.Lippincott's illustrated reviews. Philadelphia: Lippincott Williams &Wilkins; and Golan, D. E. (2008). Principles of pharmacology: thepathophysiologic basis of drug therapy. Philadelphia, Pa., [etc.]:Lippincott Williams & Wilkins.

In another embodiment of the invention, the anti-IL-6 antibodiesdescribed herein, or IL-6 binding fragments thereof, as well ascombinations of said antibody fragments, are administered to a subjectin a pharmaceutical formulation.

A “pharmaceutical composition” refers to a chemical or biologicalcomposition suitable for administration to a mammal. Such compositionsmay be specifically formulated for administration via one or more of anumber of routes, including but not limited to buccal, epicutaneous,epidural, inhalation, intraarterial, intracardial,intracerebroventricular, intradermal, intramuscular, intranasal,intraocular, intraperitoneal, intraspinal, intrathecal, intravenous,oral, parenteral, rectally via an enema or suppository, subcutaneous,subdermal, sublingual, transdermal, and transmucosal. In addition,administration can occur by means of injection, powder, liquid, gel,drops, or other means of administration.

In one embodiment of the invention, the anti-IL-6 antibodies describedherein, or IL-6 binding fragments thereof, as well as combinations ofsaid antibody fragments, may be optionally administered in combinationwith one or more active agents. Such active agents include analgesic,antipyretic, anti-inflammatory, antibiotic, antiviral, and anti-cytokineagents. Active agents include agonists, antagonists, and modulators ofTNF-α, IL-2, IL-4, IL-6, IL-10, IL-12, IL-13, IL-18, IFN-α, IFN-γ, BAFF,CXCL13, IP-10, VEGF, EPO, EGF, HRG, Hepatocyte Growth Factor (HGF),Hepcidin, including antibodies reactive against any of the foregoing,and antibodies reactive against any of their receptors. Active agentsalso include 2-Arylpropionic acids, Aceclofenac, Acemetacin,Acetylsalicylic acid (Aspirin), Alclofenac, Alminoprofen, Amoxiprin,Ampyrone, Arylalkanoic acids, Azapropazone, Benorylate/Benorilate,Benoxaprofen, Bromfenac, Carprofen, Celecoxib, Choline magnesiumsalicylate, Clofezone, COX-2 inhibitors, Dexibuprofen, Dexketoprofen,Diclofenac, Diflunisal, Droxicam, Ethenzamide, Etodolac, Etoricoxib,Faislamine, fenamic acids, Fenbufen, Fenoprofen, Flufenamic acid,Flunoxaprofen, Flurbiprofen, Ibuprofen, Ibuproxam, Indometacin,Indoprofen, Kebuzone, Ketoprofen, Ketorolac, Lornoxicam, Loxoprofen,Lumiracoxib, Magnesium salicylate, Meclofenamic acid, Mefenamic acid,Meloxicam, Metamizole, Methyl salicylate, Mofebutazone, Nabumetone,Naproxen, N-Arylanthranilic acids, Oxametacin, Oxaprozin, Oxicams,Oxyphenbutazone, Parecoxib, Phenazone, Phenylbutazone, Phenylbutazone,Piroxicam, Pirprofen, profens, Proglumetacin, Pyrazolidine derivatives,Rofecoxib, Salicyl salicylate, Salicylamide, Salicylates,Sulfinpyrazone, Sulindac, Suprofen, Tenoxicam, Tiaprofenic acid,Tolfenamic acid, Tolmetin, and Valdecoxib. Antibiotics include Amikacin,Aminoglycosides, Amoxicillin, Ampicillin, Ansamycins, Arsphenamine,Azithromycin, Azlocillin, Aztreonam, Bacitracin, Carbacephem,Carbapenems, Carbenicillin, Cefaclor, Cefadroxil, Cefalexin, Cefalothin,Cefalotin, Cefamandole, Cefazolin, Cefdinir, Cefditoren, Cefepime,Cefixime, Cefoperazone, Cefotaxime, Cefoxitin, Cefpodoxime, Cefprozil,Ceftazidime, Ceftibuten, Ceftizoxime, Ceftobiprole, Ceftriaxone,Cefuroxime, Cephalosporins, Chloramphenicol, Cilastatin, Ciprofloxacin,Clarithromycin, Clindamycin, Cloxacillin, Colistin, Co-trimoxazole,Dalfopristin, Demeclocycline, Dicloxacillin, Dirithromycin, Doripenem,Doxycycline, Enoxacin, Ertapenem, Erythromycin, Ethambutol,Flucloxacillin, Fosfomycin, Furazolidone, Fusidic acid, Gatifloxacin,Geldanamycin, Gentamicin, Glycopeptides, Herbimycin, Imipenem,Isoniazid, Kanamycin, Levofloxacin, Lincomycin, Linezolid, Lomefloxacin,Loracarbef, Macrolides, Mafenide, Meropenem, Meticillin, Metronidazole,Mezlocillin, Minocycline, Monobactams, Moxifloxacin, Mupirocin,Nafcillin, Neomycin, Netilmicin, Nitrofurantoin, Norfloxacin, Ofloxacin,Oxacillin, Oxytetracycline, Paromomycin, Penicillin, Penicillins,Piperacillin, Platensimycin, Polymyxin B, Polypeptides, Prontosil,Pyrazinamide, Quinolones, Quinupristin, Rifampicin, Rifampin,Roxithromycin, Spectinomycin, Streptomycin, Sulfacetamide,Sulfamethizole, Sulfanilimide, Sulfasalazine, Sulfisoxazole,Sulfonamides, Teicoplanin, Telithromycin, Tetracycline, Tetracyclines,Ticarcillin, Tinidazole, Tobramycin, Trimethoprim,Trimethoprim-Sulfamethoxazole, Troleandomycin, Trovafloxacin, andVancomycin. Active agents also include Aldosterone, Beclometasone,Betamethasone, Corticosteroids, Cortisol, Cortisone acetate,Deoxycorticosterone acetate, Dexamethasone, Fludrocortisone acetate,Glucocorticoids, Hydrocortisone, Methylprednisolone, Prednisolone,Prednisone, Steroids, and Triamcinolone. Antiviral agents includeabacavir, aciclovir, acyclovir, adefovir, amantadine, amprenavir, anantiretroviral fixed dose combination, an antiretroviral synergisticenhancer, arbidol, atazanavir, atripla, brivudine, cidofovir, combivir,darunavir, delavirdine, didanosine, docosanol, edoxudine, efavirenz,emtricitabine, enfuvirtide, entecavir, entry inhibitors, famciclovir,fomivirsen, fosamprenavir, foscarnet, fosfonet, fusion inhibitor,ganciclovir, gardasil, ibacitabine, idoxuridine, imiquimod, imunovir,indinavir, inosine, integrase inhibitor, interferon, interferon type I,interferon type II, interferon type III, lamivudine, lopinavir,loviride, maraviroc, MK-05I8, moroxydine, nelfinavir, nevirapine,nexavir, nucleoside analogues, oseltamivir, penciclovir, peramivir,pleconaril, podophyllotoxin, protease inhibitor, reverse transcriptaseinhibitor, ribavirin, rimantadine, ritonavir, saquinavir, stavudine,tenofovir, tenofovir disoproxil, tipranavir, trifluridine, trizivir,tromantadine, truvada, valaciclovir, valganciclovir, vicriviroc,vidarabine, viramidine, zalcitabine, zanamivir, and zidovudine. Anysuitable combination of these active agents is also contemplated.

A “pharmaceutical excipient” or a “pharmaceutically acceptableexcipient” is a carrier, usually a liquid, in which an activetherapeutic agent is formulated. In one embodiment of the invention, theactive therapeutic agent is a humanized antibody described herein, orone or more fragments thereof. The excipient generally does not provideany pharmacological activity to the formulation, though it may providechemical and/or biological stability, and release characteristics.Exemplary formulations can be found, for example, in Remington'sPharmaceutical Sciences, 19^(th) Ed., Grennaro, A., Ed., 1995 which isincorporated by reference.

As used herein “pharmaceutically acceptable carrier” or “excipient”includes any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents that arephysiologically compatible. In one embodiment, the carrier is suitablefor parenteral administration. Alternatively, the carrier can besuitable for intravenous, intraperitoneal, intramuscular, or sublingualadministration. Pharmaceutically acceptable carriers include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions. The use of such media and agents for pharmaceuticallyactive substances is well known in the art. Except insofar as anyconventional media or agent is incompatible with the active compound,use thereof in the pharmaceutical compositions of the invention iscontemplated. Supplementary active compounds can also be incorporatedinto the compositions.

Pharmaceutical compositions typically must be sterile and stable underthe conditions of manufacture and storage. The invention contemplatesthat the pharmaceutical composition is present in lyophilized form. Thecomposition can be formulated as a solution, microemulsion, liposome, orother ordered structure suitable to high drug concentration. The carriercan be a solvent or dispersion medium containing, for example, water,ethanol, polyol (for example, glycerol, propylene glycol, and liquidpolyethylene glycol), and suitable mixtures thereof. The inventionfurther contemplates the inclusion of a stabilizer in the pharmaceuticalcomposition.

In many cases, it will be preferable to include isotonic agents, forexample, sugars, polyalcohols such as mannitol, sorbitol, or sodiumchloride in the composition. Prolonged absorption of the injectablecompositions can be brought about by including in the composition anagent which delays absorption, for example, monostearate salts andgelatin. Moreover, the alkaline polypeptide can be formulated in a timerelease formulation, for example in a composition which includes a slowrelease polymer. The active compounds can be prepared with carriers thatwill protect the compound against rapid release, such as a controlledrelease formulation, including implants and microencapsulated deliverysystems. Biodegradable, biocompatible polymers can be used, such asethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers(PLG). Many methods for the preparation of such formulations are knownto those skilled in the art.

For each of the recited embodiments, the compounds can be administeredby a variety of dosage forms. Any biologically-acceptable dosage formknown to persons of ordinary skill in the art, and combinations thereof,are contemplated. Examples of such dosage forms include, withoutlimitation, reconstitutable powders, elixirs, liquids, solutions,suspensions, emulsions, powders, granules, particles, microparticles,dispersible granules, cachets, inhalants, aerosol inhalants, patches,particle inhalants, implants, depot implants, injectables (includingsubcutaneous, intramuscular, intravenous, and intradermal), infusions,and combinations thereof.

The above description of various illustrated embodiments of theinvention is not intended to be exhaustive or to limit the invention tothe precise form disclosed. While specific embodiments of, and examplesfor, the invention are described herein for illustrative purposes,various equivalent modifications are possible within the scope of theinvention, as those skilled in the relevant art will recognize. Theteachings provided herein of the invention can be applied to otherpurposes, other than the examples described above.

These and other changes can be made to the invention in light of theabove detailed description. In general, in the following claims, theterms used should not be construed to limit the invention to thespecific embodiments disclosed in the specification and the claims.Accordingly, the invention is not limited by the disclosure, but insteadthe scope of the invention is to be determined entirely by the followingclaims.

The invention may be practiced in ways other than those particularlydescribed in the foregoing description and examples. Numerousmodifications and variations of the invention are possible in light ofthe above teachings and, therefore, are within the scope of the appendedclaims.

Certain teachings related to methods for obtaining a clonal populationof antigen-specific B cells were disclosed in U.S. Provisional patentapplication No. 60/801,412, filed May 19, 2006, the disclosure of whichis herein incorporated by reference in its entirety.

Certain teachings related to humanization of rabbit-derived monoclonalantibodies and preferred sequence modifications to maintain antigenbinding affinity were disclosed in International application Ser. No.12/124,723, entitled “Novel Rabbit Antibody Humanization Method andHumanized Rabbit Antibodies”, filed May 21, 2008, now abandoned, thedisclosure of which is herein incorporated by reference in its entirety.

Certain teachings related to producing antibodies or fragments thereofusing mating competent yeast and corresponding methods were disclosed inU.S. patent application Ser. No. 11/429,053, filed May 8, 2006, now U.S.Pat. No. 7,927,863, the disclosure of which is herein incorporated byreference in its entirety. Each of U.S. Ser. No. 12/153,611, filed May21, 2008, now U.S. Pat. Nos. 9,056,905, 12/124,723 filed May 21, 2008,now abandoned, and U.S. Provisional Application Ser. No. 60/924,551,filed May 21, 2007, now expired, is hereby incorporated by reference inits entirety.

Certain teachings related to anti-IL-6 antibodies, methods of producingantibodies or fragments thereof using mating competent yeast andcorresponding methods were disclosed in U.S. provisional patentapplication No. 60/924,550, filed May 21, 2007, the disclosure of whichis herein incorporated by reference in its entirety.

Certain anti-IL-6 antibody polynucleotides and polypeptides aredisclosed in the sequence listing accompanying this patent applicationfiling, and the disclosure of said sequence listing is hereinincorporated by reference in its entirety.

The entire disclosure of each document cited (including patents, patentapplications, journal articles, abstracts, manuals, books, or otherdisclosures) in the Background of the Invention, Detailed Description,and Examples is herein incorporated by reference in their entireties.

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the subject invention, and are not intended to limit thescope of what is regarded as the invention. Efforts have been made toensure accuracy with respect to the numbers used (e.g. amounts,temperature, concentrations, etc.) but some experimental errors anddeviations should be allowed for. Unless otherwise indicated, parts areparts by weight, molecular weight is average molecular weight,temperature is in degrees centigrade; and pressure is at or nearatmospheric.

EXAMPLES Example 1 Production of Enriched Antigen-Specific B CellAntibody Culture

Panels of antibodies are derived by immunizing traditional antibody hostanimals to exploit the native immune response to a target antigen ofinterest. Typically, the host used for immunization is a rabbit or otherhost that produces antibodies using a similar maturation process andprovides for a population of antigen-specific B cells producingantibodies of comparable diversity, e.g., epitopic diversity. Theinitial antigen immunization can be conducted using complete Freund'sadjuvant (CFA), and the subsequent boosts effected with incompleteadjuvant. At about 50-60 days after immunization, preferably at day 55,antibody titers are tested, and the Antibody Selection (ABS) process isinitiated if appropriate titers are established. The two key criteriafor ABS initiation are potent antigen recognition and function-modifyingactivity in the polyclonal sera.

At the time positive antibody titers are established, animals aresacrificed and B cell sources isolated. These sources include: thespleen, lymph nodes, bone marrow, and peripheral blood mononuclear cells(PBMCs). Single cell suspensions are generated, and the cell suspensionsare washed to make them compatible for low temperature long termstorage. The cells are then typically frozen.

To initiate the antibody identification process, a small fraction of thefrozen cell suspensions are thawed, washed, and placed in tissue culturemedia. These suspensions are then mixed with a biotinylated form of theantigen that was used to generate the animal immune response, andantigen-specific cells are recovered using the Miltenyi magnetic beadcell selection methodology. Specific enrichment is conducted usingstreptavidin beads. The enriched population is recovered and progressedin the next phase of specific B cell isolation.

Example 2 Production of Clonal, Antigen-Specific B Cell-ContainingCulture

Enriched B cells produced according to Example 1 are then plated atvarying cell densities per well in a 96 well microtiter plate.Generally, this is at 50, 100, 250, or 500 cells per well with 10 platesper group. The media is supplemented with 4% activated rabbit T cellconditioned media along with 50K frozen irradiated EL4B feeder cells.These cultures are left undisturbed for 5-7 days at which timesupernatant-containing secreted antibody is collected and evaluated fortarget properties in a separate assay setting. The remaining supernatantis left intact, and the plate is frozen at −70° C. Under theseconditions, the culture process typically results in wells containing amixed cell population that comprises a clonal population ofantigen-specific B cells, i.e., a single well will only contain a singlemonoclonal antibody specific to the desired antigen.

Example 3 Screening of Antibody Supernatants for Monoclonal Antibody ofDesired Specificity and/or Functional Properties

Antibody-containing supernatants derived from the well containing aclonal antigen-specific B cell population produced according to Example2 are initially screened for antigen recognition using ELISA methods.This includes selective antigen immobilization (e.g., biotinylatedantigen capture by streptavidin coated plate), non-specific antigenplate coating, or alternatively, through an antigen build-up strategy(e.g., selective antigen capture followed by binding partner addition togenerate a heteromeric protein-antigen complex). Antigen-positive wellsupernatants are then optionally tested in a function-modifying assaythat is strictly dependant on the ligand. One such example is an invitro protein-protein interaction assay that recreates the naturalinteraction of the antigen ligand with recombinant receptor protein.Alternatively, a cell-based response that is ligand dependent and easilymonitored (e.g., proliferation response) is utilized. Supernatant thatdisplays significant antigen recognition and potency is deemed apositive well. Cells derived from the original positive well are thentransitioned to the antibody recovery phase.

Example 4 Recovery of Single, Antibody-Producing B Cell of DesiredAntigen Specificity

Cells are isolated from a well that contains a clonal population ofantigen-specific B cells (produced according to Example 2 or 3), whichsecrete a single antibody sequence. The isolated cells are then assayedto isolate a single, antibody-secreting cell. Dynal streptavidin beadsare coated with biotinylated target antigen under buffered medium toprepare antigen-containing microbeads compatible with cell viability.Next antigen-loaded beads, antibody-producing cells from the positivewell, and a fluorescein isothiocyanate (FITC)-labeled anti-host H&L IgGantibody (as noted, the host can be any mammalian host, e.g., rabbit,mouse, rat, etc.) are incubated together at 37° C. This mixture is thenre-pipetted in aliquots onto a glass slide such that each aliquot has onaverage a single, antibody-producing B-cell. The antigen-specific,antibody-secreting cells are then detected through fluorescencemicroscopy. Secreted antibody is locally concentrated onto the adjacentbeads due to the bound antigen and provides localization informationbased on the strong fluorescent signal. Antibody-secreting cells areidentified via FITC detection of antibody-antigen complexes formedadjacent to the secreting cell. The single cell found in the center ofthis complex is then recovered using a micromanipulator. The cell issnap-frozen in an eppendorf PCR tube for storage at −80° C. untilantibody sequence recovery is initiated.

Example 5 Isolation of Antibody Sequences from Antigen-Specific B Cell

Antibody sequences are recovered using a combined RT-PCR based methodfrom a single isolated B-cell produced according to Example 4 or anantigenic specific B cell isolated from the clonal B cell populationobtained according to Example 2. Primers are designed to anneal inconserved and constant regions of the target immunoglobulin genes (heavyand light), such as rabbit immunoglobulin sequences, and a two-stepnested PCR recovery step is used to obtain the antibody sequence.Amplicons from each well are analyzed for recovery and size integrity.The resulting fragments are then digested with AluI to fingerprint thesequence clonality. Identical sequences display a common fragmentationpattern in their electrophoretic analysis. Significantly, this commonfragmentation pattern which proves cell clonality is generally observedeven in the wells originally plated up to 1000 cells/well. The originalheavy and light chain amplicon fragments are then restriction enzymedigested with HindIII and XhoI or HindIII and BsiWI to prepare therespective pieces of DNA for cloning. The resulting digestions are thenligated into an expression vector and transformed into bacteria forplasmid propagation and production. Colonies are selected for sequencecharacterization.

Example 6 Recombinant Production of Monoclonal Antibody of DesiredAntigen Specificity and/or Functional Properties

Correct full-length antibody sequences for each well containing a singlemonoclonal antibody is established and miniprep DNA is prepared usingQiagen solid-phase methodology. This DNA is then used to transfectmammalian cells to produce recombinant full-length antibody. Crudeantibody product is tested for antigen recognition and functionalproperties to confirm the original characteristics are found in therecombinant antibody protein. Where appropriate, large-scale transientmammalian transfections are completed, and antibody is purified throughProtein A affinity chromatography. Kd is assessed using standard methods(e.g., Biacore) as well as IC50 in a potency assay.

Example 7 Preparation of Antibodies that Bind Human IL-6

By using the antibody selection protocol described herein, one cangenerate an extensive panel of antibodies. The antibodies have highaffinity towards IL-6 (single to double digit pM Kd) and demonstratepotent antagonism of IL-6 in multiple cell-based screening systems(T1165 and HepG2). Furthermore, the collection of antibodies displaydistinct modes of antagonism toward IL-6-driven processes.

Immunization Strategy

Rabbits were immunized with huIL-6 (R&R). Immunization consisted of afirst subcutaneous (sc) injection of 100 μg in complete Freund'sadjuvant (CFA) (Sigma) followed by two boosts, two weeks apart, of 50 μgeach in incomplete Freund's adjuvant (IFA) (Sigma). Animals were bled onday 55, and serum titers were determined by ELISA (antigen recognition)and by non-radioactive proliferation assay (Promega) using the T1165cell line.

Antibody Selection Titer Assessment

Antigen recognition was determined by coating Immulon 4 plates (Thermo)with 1 μg/ml of huIL-6 (50 μl/well) in phosphate buffered saline (PBS,Hyclone) overnight at 4° C. On the day of the assay, plates were washed3 times with PBS/Tween 20 (PBST tablets, Calbiochem). Plates were thenblocked with 200 μl/well of 0.5% fish skin gelatin (FSG, Sigma) in PBSfor 30 minutes at 37° C. Blocking solution was removed, and plates wereblotted. Serum samples were made (bleeds and pre-bleeds) at a startingdilution of 1:100 (all dilutions were made in FSG 50 μl/well) followedby 1:10 dilutions across the plate (column 12 was left blank forbackground control). Plates were incubated for 30 minutes at 37° C.Plates were washed 3 times with PBS/Tween 20. Goat anti-rabbit FC-HRP(Pierce) diluted 1:5000 was added to all wells (50 μl/well), and plateswere incubated for 30 minutes at 37° C. Plates were washed as describedabove. 50 μl/well of TMB-Stable stop (Fitzgerald Industries) was addedto plates, and color was allowed to develop, generally for 3 to 5minutes. The development reaction was stopped with 50 μl/well 0.5 M HCl.Plates were read at 450 nm. Optical density (OD) versus dilution wasplotted using Graph Pad Prizm software, and titers were determined.

Functional Titer Assessment

The functional activity of the samples was determined by a T1165proliferation assay. T1165 cells were routinely maintained in modifiedRPMI medium (Hyclone) supplemented with Hepes, sodium pyruvate, sodiumbicarbonate, L-glutamine, high glucose, penicillin/streptomycin, 10%heat inactivated fetal bovine serum (FBS) (all supplements fromHyclone), 2-mercaptoethanol (Sigma), and 10 ng/ml of huIL-6 (R&D). Onthe day of the assay, cell viability was determined by trypan blue(Invitrogen), and cells were seeded at a fixed density of 20,000cells/well. Prior to seeding, cells were washed twice in the mediumdescribed above without human-IL-6 (by centrifuging at 13000 rpm for 5minutes and discarding the supernatant). After the last wash, cells wereresuspended in the same medium used for washing in a volume equivalentto 50 μl/well. Cells were set aside at room temperature.

In a round-bottom, 96-well plate (Costar), serum samples were addedstarting at 1:100, followed by a 1:10 dilution across the plate (columns2 to 10) at 30 μl/well in replicates of 5 (rows B to F: dilution made inthe medium described above with no huIL-6). Column 11 was medium onlyfor IL-6 control. 30 μl/well of huIL-6 at 4× concentration of the finalEC50 (concentration previously determined) were added to all wells(huIL-6 was diluted in the medium described above). Wells were incubatedfor 1 hour at 37° C. to allow antibody binding to occur. After 1 hour,50 μl/well of antibody-antigen (Ab-Ag) complex were transferred to aflat-bottom, 96-well plate (Costar) following the plate map format laidout in the round-bottom plate. On Row G, 50 μl/well of medium were addedto all wells (columns 2 to 11) for background control. 50 μl/well of thecell suspension set aside were added to all wells (columns 2 to 11, rowsB to G). On Columns 1 and 12 and on rows A and H, 200 μl/well of mediumwas added to prevent evaporation of test wells and to minimize edgeeffect. Plates were incubated for 72 h at 37° C. in 4% CO2. At 72 h, 20l/well of CellTiter96 (Promega) reagents was added to all test wells permanufacturer protocol, and plates were incubated for 2 h at 37° C. At 2h, plates were gently mixed on an orbital shaker to disperse cells andto allow homogeneity in the test wells. Plates were read at 490 nmwavelength. Optical density (OD) versus dilution was plotted using GraphPad Prizm software, and functional titer was determined. A positiveassay control plate was conducted as described above using MAB2061 (R&DSystems) at a starting concentration of 1 μg/ml (final concentration)followed by 1:3 dilutions across the plate.

Tissue Harvesting

Once acceptable titers were established, the rabbit(s) were sacrificed.Spleen, lymph nodes, and whole blood were harvested and processed asfollows:

Spleen and lymph nodes were processed into a single cell suspension bydisassociating the tissue and pushing through sterile wire mesh at 70 μm(Fisher) with a plunger of a 20 cc syringe. Cells were collected in themodified RPMI medium described above without huIL-6, but with lowglucose. Cells were washed twice by centrifugation. After the last wash,cell density was determined by trypan blue. Cells were centrifuged at1500 rpm for 10 minutes; the supernatant was discarded. Cells wereresuspended in the appropriate volume of 10% dimethyl sulfoxide (DMSO,Sigma) in FBS (Hyclone) and dispensed at 1 ml/vial. Vials were thenstored at −70° C. for 24 h prior to being placed in a liquid nitrogen(LN2) tank for long-term storage.

Peripheral blood mononuclear cells (PBMCs) were isolated by mixing wholeblood with equal parts of the low glucose medium described above withoutFBS. 35 ml of the whole blood mixture was carefully layered onto 8 ml ofLympholyte Rabbit (Cedarlane) into a 45 ml conical tube (Corning) andcentrifuged 30 minutes at 2500 rpm at room temperature without brakes.After centrifugation, the PBMC layers were carefully removed using aglass Pasteur pipette (VWR), combined, and placed into a clean 50 mlvial. Cells were washed twice with the modified medium described aboveby centrifugation at 1500 rpm for 10 minutes at room temperature, andcell density was determined by trypan blue staining. After the lastwash, cells were resuspended in an appropriate volume of 10% DMSO/FBSmedium and frozen as described above.

B Cell Culture

On the day of setting up B cell culture, PBMC, splenocyte, or lymph nodevials were thawed for use. Vials were removed from LN2 tank and placedin a 37° C. water bath until thawed. Contents of vials were transferredinto 15 ml conical centrifuge tube (Corning) and 10 ml of modified RPMIdescribed above was slowly added to the tube. Cells were centrifuged for5 minutes at 1.5K rpm, and the supernatant was discarded. Cells wereresuspended in 10 ml of fresh media. Cell density and viability wasdetermined by trypan blue. Cells were washed again and resuspended at1E07 cells/80 ul medium. Biotinylated huIL-6 (B huIL-6) was added to thecell suspension at the final concentration of 3 ug/mL and incubated for30 minutes at 4° C. Unbound B huIL-6 was removed with two 10 ml washesof phosphate-buffered (PBF):Ca/Mg free PBS (Hyclone), 2 mMethylenediamine tetraacetic acid (EDTA), 0.5% bovine serum albumin (BSA)(Sigma-biotin free). After the second wash, cells were resuspended at1E07 cells/80 μl PBF. 20 μl of MACS® streptavidin beads (Milteni)/10E7cells were added to the cell suspension. Cells were incubated at 4° C.for 15 minutes. Cells were washed once with 2 ml of PBF/10E7 cells.After washing, the cells were resuspended at 1E08 cells/500 μl of PBFand set aside. A MACS® MS column (Milteni) was pre-rinsed with 500 ml ofPBF on a magnetic stand (Milteni). Cell suspension was applied to thecolumn through a pre-filter, and unbound fraction was collected. Thecolumn was washed with 1.5 ml of PBF buffer. The column was removed fromthe magnet stand and placed onto a clean, sterile 5 ml PolypropyleneFalcon tube. 1 ml of PBF buffer was added to the top of the column, andpositive selected cells were collected. The yield and viability ofpositive and negative cell fraction was determined by trypan bluestaining. Positive selection yielded an average of 1% of the startingcell concentration.

A pilot cell screen was established to provide information on seedinglevels for the culture. Three 10-plate groups (a total of 30 plates)were seeded at 50, 100, and 200 enriched B cells/well. In addition, eachwell contained 50K cells/well of irradiated EL-4.B5 cells (5,000 Rads)and an appropriate level of T cell supernatant (ranging from 1-5%depending on preparation) in high glucose modified RPMI medium at afinal volume of 250 μl/well. Cultures were incubated for 5 to 7 days at37° C. in 4% CO2.

Identification of Selective Antibody Secreting B Cells

Cultures were tested for antigen recognition and functional activitybetween days 5 and 7.

Antigen Recognition Screening

The ELISA format used is as described above except 50 μl of supernatantfrom the B cell cultures (BCC) wells (all 30 plates) was used as thesource of the antibody. The conditioned medium was transferred toantigen-coated plates. After positive wells were identified, thesupernatant was removed and transferred to a 96-well master plate(s).The original culture plates were then frozen by removing all thesupernatant except 40 μl/well and adding 60 μl/well of 16% DMSO in FBS.Plates were wrapped in paper towels to slow freezing and placed at −70°C.

Functional Activity Screening

Master plates were then screened for functional activity in the T1165proliferation assay as described before, except row B was media only forbackground control, row C was media+IL-6 for positive proliferationcontrol, and rows D-G and columns 2-11 were the wells from the BCC (50μl/well, single points). 40 μl of IL-6 was added to all wells except themedia row at 2.5 times the EC50 concentration determined for the assay.After 1 h incubation, the Ab/Ag complex was transferred to a tissueculture (TC) treated, 96-well, flat-bottom plate. 20 μl of cellsuspension in modified RPMI medium without huIL-6 (T1165 at 20,000cells/well) was added to all wells (100 μl final volume per well).Background was subtracted, and observed OD values were transformed into% of inhibition.

B Cell Recovery

Plates containing wells of interest were removed from −70° C., and thecells from each well were recovered with 5-200 μl washes of medium/well.The washes were pooled in a 1.5 ml sterile centrifuge tube, and cellswere pelleted for 2 minutes at 1500 rpm.

The tube was inverted, the spin repeated, and the supernatant carefullyremoved. Cells were resuspended in 100 μl/tube of medium. 100 μlbiotinylated IL-6 coated streptavidin M280 dynabeads (Invitrogen) and 16μl of goat anti-rabbit H&L IgG-FITC diluted 1:100 in medium was added tothe cell suspension.

20 μl of cell/beads/FITC suspension was removed, and 5 μl droplets wereprepared on a glass slide (Corning) previously treated with Sigmacote(Sigma), 35 to 40 droplets/slide. An impermeable barrier of parafin oil(JT Baker) was added to submerge the droplets, and the slide wasincubated for 90 minutes at 37° C., 4% CO2 in the dark.

Specific B cells that produce antibody can be identified by thefluorescent ring around them due to antibody secretion, recognition ofthe bead-associated biotinylated antigen, and subsequent detection bythe fluorescent-IgG detection reagent. Once a cell of interest wasidentified, the cell in the center of the fluorescent ring was recoveredvia a micromanipulator (Eppendorf). The single cell synthesizing andexporting the antibody was transferred into a 250 μl microcentrifugetube and placed in dry ice. After recovering all cells of interest,these were transferred to −70° C. for long-term storage.

Example 8 Yeast Cell Expression

Antibody Genes:

Genes were cloned and constructed that directed the synthesis of achimeric humanized rabbit monoclonal antibody.

Expression Vector:

The vector contains the following functional components: 1) a mutantColE1 origin of replication, which facilitates the replication of theplasmid vector in cells of the bacterium Escherichia coli; 2) abacterial Sh ble gene, which confers resistance to the antibiotic Zeocinand serves as the selectable marker for transformations of both E. coliand P. pastoris; 3) an expression cassette composed of theglyceraldehyde dehydrogenase gene (GAP gene) promoter, fused tosequences encoding the Saccharomyces cerevisiae alpha mating factor prepro secretion leader sequence, followed by sequences encoding a P.pastoris transcriptional termination signal from the P. pastoris alcoholoxidase I gene (AOX1). The Zeocin resistance marker gene provides ameans of enrichment for strains that contain multiple integrated copiesof an expression vector in a strain by selecting for transformants thatare resistant to higher levels of Zeocin.

P. pastoris strains: P. pastoris strains met1, lys3, ura3 and ade1 maybe used. Although any two complementing sets of auxotrophic strainscould be used for the construction and maintenance of diploid strains,these two strains are especially suited for this method for two reasons.First, they grow more slowly than diploid strains that are the result oftheir mating or fusion. Thus, if a small number of haploid ade1 or ura3cells remain present in a culture or arise through meiosis or othermechanism, the diploid strain should outgrow them in culture.

The second is that it is easy to monitor the sexual state of thesestrains since diploid Ade+ colonies arising from their mating are anormal white or cream color, whereas cells of any strains that arehaploid ade1 mutants will form a colony with a distinct pink color. Inaddition, any strains that are haploid ura3 mutants are resistant to thedrug 5-fluoro-orotic acid (FOA) and can be sensitively identified byplating samples of a culture on minimal medium+ uracil plates with FOA.On these plates, only uracil-requiring ura3 mutant (presumably haploid)strains can grow and form colonies. Thus, with haploid parent strainsmarked with ade1 and ura3, one can readily monitor the sexual state ofthe resulting antibody-producing diploid strains (haploid versusdiploid).

Methods

Construction of pGAPZ-Alpha Expression Vectors for Transcription ofLight and Heavy Chain Antibody Genes.

The humanized light and heavy chain fragments were cloned into the pGAPZexpression vectors through a PCR directed process. The recoveredhumanized constructs were subjected to amplification under standard KODpolymerase (Novagen) kit conditions ((I) 94° C., 2 minutes; (2) 94° C.,30 seconds (3) 55° C., 30 seconds; (4) 72° C., 30 seconds-cyclingthrough steps 2-4 for 35 times; (5) 72° C. 2 minutes) employing thefollowing primers (I) light chain forward

AGCGCT TATTCCGCTATCCAGATGACCCAGTC(SEQ ID NO:650) the AfeI site is single underlined. The end of the HSAsignal sequence is double underlined, followed by the sequence for themature variable light chain (not underlined); the reverseCGTACGTTTGATTTCCACCTTG (SEQ ID NO:651).

Variable light chain reverse primer. BsiWI site is underlined, followedby the reverse complement for the 3′ end of the variable light chain.Upon restriction enzyme digest with AfeI and BsiWI this enable insertionin-frame with the pGAPZ vector using the human HAS leader sequence inframe with the human kapp light chain constant region for export. (2) Asimilar strategy is performed for the heavy chain. The forward primeremployed is

AGCGCT TATTCCGAGGTGCAGCTGGTGGAGTC.(SEQ ID NO:652) The AfeI site is single underlined. The end of the HSAsignal sequence is double underlined, followed by the sequence for themature variable heavy chain (not underlined). The reverse heavy chainprimer is CTCGAGACGGTGACGAGGGT (SEQ ID NO:653). The XhoI site isunderlined, followed by the reverse complement for the 3′ end of thevariable heavy chain. This enables cloning of the heavy chain in-framewith IgG-γI CHI-CH2-CH3 region previous inserted within pGAPZ using acomparable directional cloning strategy.

Transformation of Expression Vectors into Haploid ade1 ura3, met1 andlys3 Host Strains of P. pastoris.

All methods used for transformation of haploid P. pastoris strains andgenetic manipulation of the P. pastoris sexual cycle are as described inHiggins, D. R., and Cregg, J. M., Eds. 1998. Pichia Protocols. Methodsin Molecular Biology. Humana Press, Totowa, N.J.

Prior to transformation, each expression vector is linearized within theGAP promoter sequences with AvrII to direct the integration of thevectors into the GAP promoter locus of the P. pastoris genome. Samplesof each vector are then individually transformed into electrocompetentcultures of the ade1, ura3, met1 and lys3 strains by electroporation andsuccessful transformants are selected on YPD Zeocin plates by theirresistance to this antibiotic. Resulting colonies are selected, streakedfor single colonies on YPD Zeocin plates and then examined for thepresence of the antibody gene insert by a PCR assay on genomic DNAextracted from each strain for the proper antibody gene insert and/or bythe ability of each strain to synthesize an antibody chain by a colonylift/immunoblot method (Wung et al. Biotechniques 21 808-812 (1996).Haploid ade1, met1 and lys3 strains expressing one of the three heavychain constructs are collected for diploid constructions along withhaploid ura3 strain expressing light chain gene. The haploid expressingheavy chain genes are mated with the appropriate light chain haploidura3 to generate diploid secreting protein.

Mating of haploid strains synthesizing a single antibody chain andselection of diploid derivatives synthesizing tetrameric functionalantibodies. To mate P. pastoris haploid strains, each ade1 (or met1 orlys3) heavy chain producing strain to be crossed is streaked across arich YPD plate and the ura3 light chain producing strain is streakedacross a second YPD plate (˜10 streaks per plate). After one or two daysincubation at 30° C., cells from one plate containing heavy chainstrains and one plate containing ura3 light chain strains aretransferred to a sterile velvet cloth on a replica-plating block in across hatched pattern so that each heavy chain strain contain a patch ofcells mixed with each light chain strain. The cross-streaked replicaplated cells are then transferred to a mating plate and incubated at 25°C. to stimulate the initiation of mating between strains. After twodays, the cells on the mating plates are transferred again to a sterilevelvet on a replica-plating block and then transferred to minimal mediumplates. These plates are incubated at 30° C. for three days to allow forthe selective growth of colonies of prototrophic diploid strains.Colonies that arose are picked and streaked onto a second minimal mediumplate to single colony isolate and purify each diploid strain. Theresulting diploid cell lines are then examined for antibody production.

Putative diploid strains are tested to demonstrate that they are diploidand contain both expression vectors for antibody production. Fordiploidy, samples of a strain are spread on mating plates to stimulatethem to go through meiosis and form spores. Haploid spore products arecollected and tested for phenotype. If a significant percentage of theresulting spore products are single or double auxotrophs it may beconcluded that the original strain must have been diploid. Diploidstrains are examined for the presence of both antibody genes byextracting genomic DNA from each and utilizing this DNA in PCR reactionsspecific for each gene.

Fusion of haploid strains synthesizing a single antibody chain andselection of diploid derivatives synthesizing tetrameric functionalantibodies. As an alternative to the mating procedure described above,individual cultures of single-chain antibody producing haploid ade1 andura3 strains are spheroplasted and their resulting spheroplasts fusedusing polyethylene glycol/CaCl₂. The fused haploid strains are thenembedded in agar containing 1 M sorbitol and minimal medium to allowdiploid strains to regenerate their cell wall and grow into visiblecolonies. Resulting colonies are picked from the agar, streaked onto aminimal medium plate, and the plates are incubated for two days at 30°C. to generate colonies from single cells of diploid cell lines. Theresulting putative diploid cell lines are then examined for diploidy andantibody production as described above.

Purification and analysis of antibodies. A diploid strain for theproduction of full length antibody is derived through the mating of met1light chain and lys3 heavy chain using the methods described above.Culture media from shake-flask or fermenter cultures of diploid P.pastoris expression strains are collected and examined for the presenceof antibody protein via SDS-PAGE and immunoblotting using antibodiesdirected against heavy and light chains of human IgG, or specificallyagainst the heavy chain of IgG.

To purify the yeast secreted antibodies, clarified media from antibodyproducing cultures are passed through a protein A column and afterwashing with 20 mM sodium phosphate, pH 7.0, binding buffer, protein Abound protein is eluted using 0.1 M glycine HCl buffer, pH 3.0.Fractions containing the most total protein are examined by Coomasieblue strained SDS-PAGE and immunoblotting for antibody protein. Antibodyis characterized using the ELISA described above for IL-6 recognition.

Assay for Antibody Activity.

The recombinant yeast-derived humanized antibody is evaluated forfunctional activity through the IL-6 driven T1165 cell proliferationassay and IL-6 stimulated HepG2 haptoglobin assay described above.

Example 9 Acute Phase Response Neutralization by IntravenousAdministration of Anti-IL-6 Antibody Ab1

Human IL-6 can provoke an acute phase response in rats, and one of themajor acute phase proteins that is stimulated in the rat is α-2macroglobulin (A2M). A study was designed to assess the dose of antibodyAb1 required to ablate the A2M response to a single s.c. injection of100 μg of human IL-6 given one hour after different doses (0.03, 0.1,0.3, 1, and 3 mg/kg) of antibody Ab1 administered intravenously (n=10rats/dose level) or polyclonal human IgG1 as the control (n=10 rats).Plasma was recovered and the A2M was quantitated via a commercialsandwich ELISA kit (ICL Inc., Newberg Oreg.; cat. no.-E-25A2M). Theendpoint was the difference in the plasma concentration of A2M at the 24hour time point (post-Ab1). The results are presented in FIG. 4.

The ID50 for antibody Ab1 was 0.1 mg/kg with complete suppression of theA2M response at the 0.3 mg/kg. This firmly establishes in vivoneutralization of human IL-6 can be accomplished by antibody Ab1.

Example 10 RXF393 Cachexia Model Study 1 Introduction

The human renal cell cancer cell line, RXF393 produces profound weightloss when transplanted into athymic nude mice. Weight loss begins aroundday 15 after transplantation with 80% of all animals losing at least 30%of their total body weight by day 18-20 after transplantation. RXF393secretes human IL-6 and the plasma concentration of human IL-6 in theseanimals is very high at around 10 ng/ml. Human IL-6 can bind murinesoluble IL-6 receptor and activate IL-6 responses in the mouse. HumanIL-6 is approximately 10 times less potent than murine IL-6 atactivating IL-6 responses in the mouse. The objectives of this studywere to determine the effect of antibody Ab1, on survival, body weight,serum amyloid A protein, hematology parameters, and tumor growth inathymic nude mice transplanted with the human renal cell cancer cellline, RXF393.

Methods

Eighty, 6 week old, male athymic nude mice were implanted with RXF393tumor fragments (30-40 mg) subcutaneously in the right flank. Animalswere then divided into eight groups of ten mice. Three groups were giveneither antibody Ab1 at 3 mg/kg, 10 mg/kg, or 30 mg/kg intravenouslyweekly on day 1, day 8, day 15 and day 22 after transplantation(progression groups). Another three groups were given either antibodyAb1 at 3 mg/kg, or 10 mg/kg, or 30 mg/kg intravenously weekly on day 8,day 15 and day 22 after transplantation (regression groups). Finally,one control group was given polyclonal human IgG 30 mg/kg and a secondcontrol group was given phosphate buffered saline intravenously weeklyon day 1, day 8, day 15 and day 22 after transplantation.

Animals were euthanized at either day 28, when the tumor reached 4,000mm³ or if they became debilitated (>30% loss of body weight). Animalswere weighed on days 1, 6 and then daily from days 9 to 28 aftertransplantation. Mean Percent Body Weight (MPBW) was used as the primaryparameter to monitor weight loss during the study. It was calculated asfollows: (Body Weight−Tumor Weight)/Baseline Body Weight×100. Tumorweight was measured on days 1, 6, 9, 12, 15, 18, 22, 25 and 28 aftertransplantation. Blood was taken under anesthesia from five mice in eachgroup on days 5 and 13 and all ten mice in each group when euthanized(day 28 in most cases). Blood was analyzed for hematology and serumamyloid A protein (SAA) concentration. An additional group of 10non-tumor bearing 6 week old, athymic nude male mice had blood samplestaken for hematology and SAA concentration estimation to act as abaseline set of values.

Results—Survival

No animals were euthanized or died in any of the antibody Ab1 groupsprior to the study termination date of day 28. In the two controlgroups, 15 animals (7/9 in the polyclonal human IgG group and 8/10 inthe phosphate buffered saline group) were found dead or were euthanizedbecause they were very debilitated (>30% loss of body weight). Mediansurvival time in both control groups was 20 days.

The survival curves for the two control groups and the antibody Ab1progression (dosed from day 1 of the study) groups are presented in FIG.5.

The survival curves for the two control groups and the antibody Ab1regression (dosed from day 8 of the study) groups are presented in FIG.6.

There was a statistically significant difference between the survivalcurves for the polyclonal human IgG (p=0.0038) and phosphate bufferedsaline (p=0.0003) control groups and the survival curve for the sixantibody Ab1 groups. There was no statistically significant differencebetween the two control groups (p=0.97).

Results—Tumor Size

Tumor size in surviving mice was estimated by palpation. For the first15 days of the study, none of the mice in any group were found dead orwere euthenized, and so comparison of tumor sizes between groups onthese days was free from sampling bias. No difference in tumor size wasobserved between the antibody Ab1 progression or regression groups andthe control groups through day 15. Comparison of the tumor size betweensurviving mice in the control and treatment groups subsequent to theonset of mortality in the controls (on day 15) was not undertakenbecause tumor size the surviving control mice was presumed to be biasedand accordingly the results of such comparison would not be meaningful.

As administration of antibody Ab1 promoted survival without any apparentreduction in tumor size, elevated serum IL-6 may contribute to mortalitythrough mechanisms independent of tumor growth. These observationssupports the hypothesis that antibody Ab1 can promote cancer patientsurvivability without directly affecting tumor growth, possibly byenhancing general patient well-being.

Results—Weight Loss

Mean Percent Body Weight (MPBW) (±SEM) versus time is shown in FIG. 27.Compared to controls, mice dosed with Ab1 were protected from weightloss. On day 18, MPBW in control mice was 75%, corresponding to anaverage weight loss of 25%. In contrast, on the same day, MPBW in Ab-1treatment groups was minimally changed (between 97% and 103%). There wasa statistically significant difference between the MPBW curves for thecontrols (receiving polyclonal human IgG or PBS) and the 10 mg/kg dosagegroup (p<0.0001) or 3 mg/kg and 30 mg/kg dosage groups (p<0.0005). Therewas no statistically significant difference between the two controlgroups.

Representative photographs of control and Ab1-treated mice (FIG. 28)illustrate the emaciated condition of the control mice, compared to thenormal appearance of the Ab1-treated mouse, at the end of the study(note externally visible tumor sites in right flank).

These results suggest that Ab1 may be useful to prevent or treatcachexia caused by elevated IL-6 in humans.

Results—Plasma Serum Amyloid A

The mean (±SEM) plasma serum amyloid A concentration versus time for thetwo control groups and the antibody Ab1 progression (dosed from day 1 ofthe study) and regression (dosed from day 8 of the study) groups arepresented in Table 1 and graphically in FIG. 32.

TABLE 1 Mean Plasma SAA - antibody Ab1, all groups versus control groupsMean Plasma Mean Plasma Mean Plasma SAA ± SEM SAA ± SEM Day 5 SAA ± SEMDay 13 Terminal Bleed (μg/ml) (μg/ml) (μg/ml) Polyclonal IgG iv 675 ±240 (n = 5) 3198 ± 628 (n = 4) 13371 ± 2413 (n = 4)   weekly from day 1PBS iv weekly 355 ± 207 (n = 5) 4844 ± 1126 (n = 5)  15826 ± 802 (n =3)   from day 1 Ab1 30 mg/kg iv 246 ± 100 (n = 5) 2979 ± 170 (n = 5) 841± 469 (n = 10) weekly from day 1 Ab1 10 mg/kg iv 3629 ± 624 (n = 5) 3096 ± 690 (n = 5) 996 ± 348 (n = 10) weekly from day 1 Ab1 3 mg/kg iv 106 ± 9 (n = 5) 1623 ± 595 (n = 4) 435 ± 70 (n = 9) weekly from day 1Ab1 30 mg/kg iv 375 ± 177 (n = 5) 1492 ± 418 (n = 4) 498 ± 83 (n = 9)weekly from day 8 Ab1 10 mg/kg iv 487 ± 170 (n = 5) 1403 ± 187 (n = 5) 396 ± 58 (n = 10) weekly from day 8 Ab1 3 mg/kg iv 1255 ± 516 (n = 5)  466 ± 157 (n = 5) 685 ± 350 (n = 5)  weekly from day 8

SAA is up-regulated via the stimulation of hIL-6 and this response isdirectly correlated with circulating levels of hIL-6 derived from theimplanted tumor. The surrogate marker provides an indirect readout foractive hIL-6. Thus in the two treatment groups described above there aresignificantly decreased levels of SAA due to the neutralization oftumor-derived hIL-6. This further supports the contention that antibodyAb1 displays in vivo efficacy.

Example 11 RXF393 Cachexia Model Study 2 Introduction

A second study was performed in the RXF-393 cachexia model wheretreatment with antibody Ab1 was started at a later stage (days 10 and 13post-transplantation) and with a more prolonged treatment phase (out to49 days post transplantation). The dosing interval with antibody Ab1 wasshortened to 3 days from 7 and also daily food consumption was measured.There was also an attempt to standardize the tumor sizes at the time ofinitiating dosing with antibody Ab1.

Methods

Eighty, 6 week old, male athymic nude mice were implanted with RXF393tumor fragments (30-40 mg) subcutaneously in the right flank. 20 micewere selected whose tumors had reached between 270-320 mg in size anddivided into two groups. One group received antibody Ab1 at 10 mg/kgi.v. every three days and the other group received polyclonal human IgG10 mg/kg every 3 days from that time-point (day 10 aftertransplantation). Another 20 mice were selected when their tumor sizehad reached 400-527 mg in size and divided into two groups. One groupreceived antibody Ab1 at 10 mg/kg i.v. every three days and the othergroup received polyclonal human IgG 10 mg/kg every 3 days from thattime-point (day 13 after transplantation). The remaining 40 mice took nofurther part in the study and were euthanized at either day 49, when thetumor reached 4,000 mm³ or if they became very debilitated (>30% loss ofbody weight).

Animals were weighed every 3-4 days from day 1 to day 49 aftertransplantation. Mean Percent Body Weight (MPBW) was used as the primaryparameter to monitor weight loss during the study. It was calculated asfollows: ((Body Weight−Tumor Weight)/Baseline Body Weight)×100. Tumorweight was measured every 3-4 days from day 5 to day 49 aftertransplantation. Food consumption was measured (amount consumed in 24hours by weight (g) by each treatment group) every day from day 10 forthe 270-320 mg tumor groups and day 13 for the 400-527 mg tumor groups.

Results—Survival

The survival curves for antibody Ab1 at 10 mg/kg i.v. every three days(270-320 mg tumor size) and for the polyclonal human IgG 10 mg/kg i.v.every three days (270-320 mg tumor size) are presented in FIG. 7.

Median survival for the antibody Ab1 at 10 mg/kg i.v. every three days(270-320 mg tumor size) was 46 days and for the polyclonal human IgG at10 mg/kg i.v. every three days (270-320 mg tumor size) was 32.5 days(p=0.0071).

The survival curves for the antibody Ab1 at 10 mg/kg i.v. every threedays (400-527 mg tumor size) and for the polyclonal human IgG at 10mg/kg i.v. every three days (400-527 mg tumor size) are presented inFIG. 8. Median survival for the antibody Ab1 at 10 mg/kg i.v. everythree days (400-527 mg tumor size) was 46.5 days and for the polyclonalhuman IgG at 10 mg/kg i.v. every three days (400-527 mg tumor size) was27 days (p=0.0481).

Example 12 Multi-Dose Pharmacokinetic Evaluation of Antibody Ab1 inNon-Human Primates

Antibody Ab1 was dosed in a single bolus infusion to a single male andsingle female cynomologus monkey in phosphate buffered saline. Plasmasamples were removed at fixed time intervals and the level of antibodyAb1 was quantitated through of the use of an antigen capture ELISAassay. Biotinylated IL-6 (50 μl of 3 μg/mL) was captured on Streptavidincoated 96 well microtiter plates. The plates were washed and blockedwith 0.5% Fish skin gelatin. Appropriately diluted plasma samples wereadded and incubated for 1 hour at room temperature. The supernatantsremoved and an anti-hFc-HRP conjugated secondary antibody applied andleft at room temperature.

The plates were then aspirated and TMB added to visualize the amount ofantibody. The specific levels were then determined through the use of astandard curve. A second dose of antibody Ab1 was administered at day 35to the same two cynomologus monkeys and the experiment replicated usingan identical sampling plan. The resulting concentrations are then plotvs. time as show in FIG. 9.

This humanized full length aglycosylated antibody expressed and purifiedPichia pastoris displays comparable characteristics to mammalianexpressed protein. In addition, multiple doses of this product displayreproducible half-lives inferring that this production platform does notgenerate products that display enhanced immunogenicity.

Example 13 Octet Mechanistic Characterization of Antibody Proteins

IL-6 signaling is dependent upon interactions between IL-6 and tworeceptors, IL-6RI (CDI26) and gp130 (IL-6 signal transducer). Todetermine the antibody mechanism of action, mechanistic studies wereperformed using bio-layer interferometry with an Octet QK instrument(ForteBio; Menlo Park, Calif.). Studies were performed in two differentconfigurations. In the first orientation, biotinylated IL-6 (R&D systemspart number 206-IL-00IMG/CF, biotinylated using Pierce EZ-linksulfo-NHS-LC-LC-biotin product number 21338 according to manufacturer'sprotocols) was initially bound to a streptavidin coated biosensor(ForteBio part number 18-5006). Binding is monitored as an increase insignal.

The IL-6 bound to the sensor was then incubated either with the antibodyin question or diluent solution alone. The sensor was then incubatedwith soluble IL-6RI (R&D systems product number 227-SR-025/CF) molecule.If the IL-6RI molecule failed to bind, the antibody was deemed to blockIL-6/IL-6RI interactions. These complexes were incubated with gpI30 (R&Dsystems 228-GP-0I0/CF) in the presence of IL-6RI for stability purposes.IfgpI30 did not bind, it was concluded that the antibody blocked gpI30interactions with IL-6.

In the second orientation, the antibody was bound to a biosensor coatedwith an anti-human IgGI Fc-specific reagent (ForteBio part numberI8-500I). The IL-6 was bound to the immobilized antibody and the sensorwas incubated with IL-6R1. If the IL-6RI did not interact with the IL-6,then it was concluded that the IL-6 binding antibody blocked IL-6/IL-6RIinteractions. In those situations where antibody/IL-6/IL-6RI wasobserved, the complex was incubated with gpI30 in the presence ofIL-6RI. If gpI30 did not interact, then it was concluded that theantibody blocked IL-6/gpI30 interactions. All studies were performed ina 200 μL final volume, at 30 C and 1000 rpms. For these studies, allproteins were diluted using ForteBio's sample diluent buffer (partnumber I8-5028).

Results are presented in FIG. 100 (A-E) and FIG. 1I.

Example 14 Peptide Mapping

In order to determine the epitope recognized by Ab1 on human IL-6, theantibody was employed in a western-blot based assay. The form of humanIL-6 utilized in this example had a sequence of 183 amino acids inlength (shown below). A 57-member library of overlapping 15 amino acidpeptides encompassing this sequence was commercially synthesized andcovalently bound to a PepSpots nitrocellulose membrane (JPT Peptidetechnologies, Berlin, Germany). The sequences of the overlapping 15amino acid peptides is shown in FIG. 12. Blots were prepared and probedaccording to the manufacturer's recommendations.

Briefly, blots were pre-wet in methanol, rinsed in PBS, and blocked forover 2 hours in 10% non-fat milk in PBS/0.05% Tween (Blocking Solution).The Ab1 antibody was used at 1 mg/ml final dilution, and theHRP-conjugated Mouse Anti-Human-Kappa secondary antibody (SouthernBioTech #9220-05) was used at a 1:5000 dilution. Antibodydilutions/incubations were performed in blocking solution. Blots weredeveloped using Amersham ECL advance reagents (GE # RPN2135) andchemiluminescent signal documented using a CCD camera (Alphalnnotec).The results of the blots is shown in FIG. 13 and FIG. 14.

The sequence of the form of human IL-6 utilized to generate peptidelibrary is set forth:

(SEQ ID NO: 1) VPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDGISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWLQDMTTHLILRSFKEFLQSSLRALRQM.

Example 15 Ab1 has High Affinity for IL-6

Surface plasmon resonance was used to measure association rate (K_(a)),dissociation rate (K_(d)) and dissociation constant (K_(D)) for Ab1 toIL-6 from rat, mouse, dog, human, and cynomolgus monkey at 25° C. (FIG.15A). The dissociation constant for human IL-6 was 4 pM, indicating veryhigh affinity. As expected, affinity generally decreased withphylogenetic distance from human. The dissociation constants of Ab1 forIL-6 of cynomolgus monkey, rat, and mouse were 31 pM, 1.4 nM, and 0.4nM, respectively. Ab1 affinity for dog IL-6 below the limit ofquantitation of the experiment.

The high affinity of Ab1 for mouse, rat, and cynomolgus monkey IL-6suggest that Ab1 may be used to inhibit IL-6 of these species. Thishypothesis was tested using a cell proliferation assay. In brief, eachspecies's IL-6 was used to stimulate proliferation of T1165 cells, andthe concentration at which Ab1 could inhibit 50% of proliferation (IC50)was measured. Inhibition was consistent with the measured dissociationconstants (FIG. 15B). These results demonstrate that Ab1 can inhibit thenative IL-6 of these species, and suggest the use of these organisms forin vitro or in vivo modeling of IL-6 inhibition by Ab1.

Example 16 Multi-Dose Pharmacokinetic Evaluation of Antibody Ab1 inHealthy Human Volunteers

Antibody Ab1 was dosed in a single bolus infusion in histidine andsorbitol to healthy human volunteers. Dosages of 1 mg, 3 mg, 10 mg, 30mg or 100 mg were administered to each individual in dosage groupscontaining five to six individuals. Plasma samples were removed at fixedtime intervals for up to twelve weeks. Human plasma was collected viavenipuncture into a vacuum collection tube containing EDTA. Plasma wasseparated and used to assess the circulating levels of Ab1 using amonoclonal antibody specific for Ab1, as follows. A 96 well microtiterplate was coated overnight with the monoclonal antibody specific for Ab1in 1×PBS overnight at 4° C. The remaining steps were conducted at roomtemperature. The wells were aspirated and subsequently blocked using0.5% Fish Skin Gelatin (FSG) (Sigma) in 1×PBS for 60 minutes. Humanplasma samples were then added and incubated for 60 minutes, thenaspirated, then 50 ul of 1 μg/mL biotinylated IL-6 was then added toeach well and incubated for 60 minutes. The wells were aspirated, and 50μL streptavidin-HRP (Pharmingen), diluted 1:5,000 in 0.5% FSG/PBS, wasadded and incubated for 45 minutes. Development was conducted usingstandard methods employing TMB for detection. Levels were thendetermined via comparison to a standard curve prepared in a comparableformat.

Average plasma concentration of Ab1 for each dosage group versus time isshown in FIG. 16. Mean AUC and C_(max) increased linearly with dosage(FIG. 17 and FIG. 18, respectively). For dosages of 30 mg and above, theaverage Ab1 half-life in each dosage group was between approximately 25and 30 days (FIG. 19).

Example 17 Pharmacokinetics of Ab1 in Patients with Advanced Cancer

Antibody Ab1 was dosed in a single bolus infusion in phosphate bufferedsaline to five individuals with advanced cancer. Each individualreceived a dosage of 80 mg (n=2) or 160 mg (n=3) of Ab1. Plasma sampleswere drawn weekly, and the level of antibody Ab1 was quantitated as inExample 16.

Average plasma concentration of Ab1 in these individuals as a functionof time is shown in FIG. 20. The average Ab1 half-life was approximately31 days.

Example 18 Unprecedented Half-Life of Ab1

Overall, the average half-life of Ab1 was approximately 31 days inhumans (for dosages of 10 mg and above), and approximately 15-21 days incynomolgus monkey. The Ab1 half-life in humans and cynomolgus monkeysare unprecedented when compared with the half-lives of other anti-IL-6antibodies (FIG. 21). As described above, Ab1 was derived fromhumanization of a rabbit antibody, and is produced from Pichia pastorisin an aglycosylated form. These characteristics results in an antibodywith very low immunogenicity in humans. Moreover, the lack ofglycosylation prevents Ab1 from interacting with the Fc receptor orcomplement. Without intent to be limited by theory, it is believed thatthe unprecedent half-life of Ab1 is at least partially attributable tothe humanization and lack of glycosylation. The particular sequenceand/or structure of the antigen binding surfaces may also contribute toAb1's half-life.

Example 19 Ab1 Effect on Hemoglobin Concentration, Plasma LipidConcentration, and Neutrophil Counts in Patients with Advanced Cancer

Antibody Ab1 was dosed in a single bolus infusion in phosphate bufferedsaline to eight individuals with advanced cancer (NSCLC, colorectalcancer, cholangiocarcinoma, or mesothelioma). Each individual received adosage of 80 mg, 160 mg, or 320 mg of Ab1. Blood samples were removedjust prior to infusion and at fixed time intervals for six weeks, andthe hemoglobin concentration, plasma lipid concentration, and neutrophilcounts were determined. Average hemoglobin concentration rose slightly(FIG. 22), as did total cholesterol and triglycerides (FIG. 23), whilemean neutrophil counts fell slightly (FIG. 24).

These results further demonstrate some of the beneficial effects ofadministration of Ab1 to chronically ill individuals. Because IL-6 isthe main cytokine resposible for the anemia of chronic disease(including cancer-related anemia), neutralization of IL-6 by Ab1increases hemoglobin concentration in these individuals. Similarly, asIL-6 is centrally important in increasing neutrophil counts ininflammation, the observed slight reduction in neutrophil counts furtherconfirms that Ab1 inhibits IL-6. Finally, IL-6 causes anorexia as wellas cachexia in these patients; neutralization of IL-6 by Ab1 results inthe return of appetite and reversal of cachexia. The increase in plasmalipid concentrations reflect the improved nutritional status of thepatients. Taken together, these results further demonstrate that Ab1effectively reverses these adverse consequences of IL-6 in thesepatients.

Example 20 Ab1 Suppresses Serum CRP in Healthy Volunteers and inPatients with Advanced Cancer Introduction

Serum CRP concentrations have been identified as a strong prognosticindicator in patients with certain forms of cancer. For example,Hashimoto et al. performed univariate and multivariate analysis ofpreoperative serum CRP concentrations in patients with hepatocellularcarcinoma in order to identify factors affecting survival and diseaserecurrence (Hashimoto, K., et al., Cancer, 103(9):1856-1864 (2005)).Patients were classified into two groups, those with serum CRPlevels>1.0 mg/dl (“the CRP positive group”) and those with serum CRPlevels<1.0 mg/dl (“the CRP negative group”). The authors identified “asignificant correlation between preoperative serum CRP level and tumorsize.” Id. Furthermore, the authors found that “[t]he overall survivaland recurrence-free survival rates in the CRP-positive group weresignificantly lower compared with the rates in the CRP-negative group.”Id. The authors concluded that the preoperative CRP level of patients isan independent and significant predictive indicator or poor prognosisand early recurrence in patients with hepatocellular carcinoma.

Similar correlations have been identified by other investigators. Forexample, Karakiewicz et al. determined that serum CRP was an independentand informative predictor of renal cell carcinoma-specific mortality(Karakiewicz, P. I., et al., Cancer, 110(6): 1241-1247 (2007)).Accordingly, there remains a need in the art for methods and/ortreatments that reduce serum C-Reactive Protein (CRP) concentrations incancer patients, and particularly those with advanced cancers.

Methods

Healthy volunteers received a single 1-hour intravenous (IV) infusion ofeither 100 mgs (5 patients), 30 mgs (5 patients), 10 mgs (6 patients), 3mgs (6 patients) or 1 mg (6 patients) of the Ab1 monoclonal antibody,while another 14 healthy volunteers received intravenous placebo.Comparatively, 2 patients with advanced forms of colorectal cancerreceived a single 1-hour intravenous (IV) infusion of 80 mgs of the Ab1monoclonal antibody. No further dosages of the Ab1 monoclonal antibodywere administered to the test population.

Patients were evaluated prior to administration of the dosage, andthereafter on a weekly basis for at least 5 weeks post dose. At the timeof each evaluation, patients were screened for serum CRP concentration.

Results

Healthy Volunteers

As noted above, serum CRP levels are a marker of inflammation;accordingly, baseline CRP levels are typically low in healthyindividuals. The low baseline CRP levels can make a further reduction inCRP levels difficult to detect. Nonetheless, a substantial reduction inserum CRP concentrations was detectable in healthy volunteers receivingall concentrations of the Ab1 monoclonal antibody, compared to controls(FIG. 25). The reduction in serum CRP levels was rapid, occurring withinone week of antibody administration, and prolonged, continuing at leastthrough the final measurement was taken (8 or 12 weeks from antibodyadministration).

Cancer Patients

Five advanced cancer patients (colorectal cancer, cholangiocarcinoma, orNSCLC) having elevated serum CRP levels were dosed with 80 mg or 160 mgof Ab1. Serum CRP levels were greatly reduced in these patients (FIG.26A). The reduction in serum CRP levels was rapid, with 90% of thedecrease occuring within one week of Ab1 administration, and prolonged,continuing at least until the final measurement was taken (up to twelveweeks). The CRP levels of two representative individuals are shown inFIG. 26B. In those individuals, the CRP levels were lowered to below thenormal reference range (less than 5-6 mg/1) within one week. Thus,administration of Ab1 to advanced cancer patients can cause a rapid andsustained suppression of serum CRP levels.

Example 21 Ab1 Improved Muscular Strength, Improved Weight, and ReducedFatigue in Patients with Advanced Cancer Introduction

Weight loss and fatigue (and accompanying muscular weakness) are verycommon symptoms of patients with advanced forms of cancer, and thesesymptoms can worsen as the cancer continues to progress. Fatigue, weightloss and muscular weakness can have significant negative effects on therecovery of patients with advanced forms of cancer, for example bydisrupting lifestyles and relationships and affecting the willingness orability of patients to continue cancer treatments. Known methods ofaddressing fatigue, weight loss and muscular weakness include regularroutines of fitness and exercise, methods of conserving the patient'senergy, and treatments that address anemia-induced fatigue and muscularweakness. Nevertheless, there remains a need in the art for methodsand/or treatments that improve fatigue, weight loss and muscularweakness in cancer patients.

Methods

Four patients with advanced forms of cancer (colorectal cancer (2),NSCLC (1), cholangiocarcinoma (1) received a single 1-hour intravenous(IV) infusion of either 80 mgs or 160 mgs of the Ab1 monoclonalantibody. No further dosages of the Ab1 monoclonal antibody wereadministered to the test population.

Patients were evaluated prior to administration of the dosage, andthereafter for at least 6 weeks post dose. At the time of eachevaluation, patients were screened for the following: a.) any change inweight; b.) fatigue as measured using the Facit-F Fatigue Subscalequestionnaire a medically recognized test for evaluating fatigue (See,e.g., Cella, D., Lai, J. S., Chang, C. H., Peterman, A., & Slavin, M.(2002). Fatigue in cancer patients compared with fatigue in the generalpopulation. Cancer, 94(2), 528-538; Cella, D., Eton, D. T., Lai, F J-S.,Peterman, A. H & Merkel, D. E. (2002). Combining anchor and distributionbased methods to derive minimal clinically important differences on theFunctional Assessment of Cancer Therapy anemia and fatigue scales.Journal of Pain & Symptom Management, 24 (6) 547-561.); and hand-gripstrength (a medically recognized test for evaluating muscle strength,typically employing a handgrip dynamometer).

Results

Weight Change

The averaged data for both dosage concentrations (80 mgs and 160 mgs) ofthe Ab1 monoclonal antibody demonstrated an increase of about 2kilograms of weight per patient over the period of 6 weeks (FIG. 29).

Fatigue

The averaged data for both dosage concentrations (80 mgs and 160 mgs) ofthe Ab1 monoclonal antibody demonstrated an increase in the mean Facit-FFS subscale score of at least about 10 points in the patient populationover the period of 6 weeks (FIG. 30).

Hand-Grip Strength

The averaged data for both dosage concentrations (80 mgs and 160 mgs) ofthe Ab1 monoclonal antibody demonstrated an increase in the meanhand-grip strength of at least about 10 percent in the patientpopulation over the period of 6 weeks (FIG. 31).

Example 22 Ab1 For Prevention of Thrombosis

Prior studies have shown that administration of an anti-IL-6 antibodycan cause decreased platelet counts. Emilie, D. et al., Blood,84(8):2472-9 (1994); Blay et al., Int J Cancer, 72(3):424-30 (1997).These results have apparently been viewed as an indicator of potentialdanger, because further decreases in platelet counts could causecomplications such as bleeding. However, Applicants have now discernedthat inhibiting IL-6 restores a normal coagulation profile, whichApplicants predict will prevent thrombosis. Decreased platelet countsresulting from inhibition of IL-6 is not a sign of potential danger butrather reflects the beneficial restoration of normal coagulation.

The mechanism by which normal coagulation is restored is believed toresult from the interplay between IL-6 and the acute phase reaction. Inresponse to elevated IL-6 levels, as for example in a cancer patient,the liver produces acute phase proteins. These acute phase proteinsinclude coagulation factors, such as Factor II, Factor V, Factor VIII,Factor IX, Factor XI, Factor XII, F/fibrin degradation products,thrombin-antithrombin III complex, fibrinogen, plasminogen, prothrombin,and von Willebrand factor. This increase in coagulation factors may bemeasured directly, or may be inferred from functional measurements ofclotting ability. Antagonists of IL-6, such as Ab1, suppresses acutephase proteins, e.g., Serum Amyloid A (see FIG. 32 and Example 10).Applicants now predict that this suppression of acute phase proteinswill restore the normal coagulation profile, and thereby preventthrombosis. The restoration of normal coagulation may cause a slightdrop in platelet counts, but the patient will nonetheless retain normalcoagulation ability and thus will not have an increased risk ofbleeding. Such a treatment will represent a vast improvement over theavailable anticoagulation therapies whose usefulness is limited by therisk of adverse side-effects, such as major bleeding.

Applicants contemplate that the same beneficial effects of inhibitingIL-6 will be obtained regardless of the method of inhibition. Suitablemethods of inhibiting IL-6 include administration of anti-IL-6antibodies, antisense therapy, soluble IL-6 receptor, etc. eitherindividually or in combinations.

Example 23 Ab1 Increases Plasma Albumin Concentration in Patients withAdvanced Cancer Introduction

Serum albumin concentrations are recognized as predictive indicators ofsurvival and/or recovery success of cancer patients. Hypoalbumeniacorrelates strongly with poor patient performance in numerous forms ofcancer. For example, in one study no patients undergoing systemicchemotherapy for metastatic pancreatic adenocarcinoma and having serumalbumin levels less than 3.5 g/dL successfully responded to systemicchemotherapy (Fujishiro, M., et al., Hepatogastroenterology,47(36):1744-46 (2000)). The authors conclude that “[p]atients with . . .hypoalbuminemia . . . might be inappropriate candidates for systemicchemotherapy and might be treated with other experimental approaches orsupportive care.” Id.

Similarly, Senior and Maroni state that “[t]he recent appreciation thathypoalbuminemia is the most powerful predictor of mortality in end-stagerenal disease highlights the critical importance of ensuring adequateprotein intake in this patient population.” (J. R. Senior and B. J.Maroni, Am. Soc. Nutr. Sci., 129:313S-314S (1999)).

In at least one study, attempts to recitfy hypoalbuminemia in 27patients with metastatic cancer by daily intravenous albumin infusion of20 g until normal serum albumin levels (>3.5 g/dl) were achieved hadlittle success. The authors note that “[a]lbumin infusion for theadvanced stage cancer patients has limited value in clinical practice.Patients with PS 4 and hypoalbuminemia have poorer prognosis.”(Demirkazik, A., et al., Proc. Am. Soc. Clin. Oncol., 21:Abstr 2892(2002)).

Accordingly, there remains a need in the art for methods and/ortreatments that improve serum albumin concentrations in cancer patientsand address hypoalbuminemic states in cancer patients, particularlythose with advanced cancers.

Methods

Four patients with advanced forms of cancer (colorectal cancer (2),NSCLC (1), cholangiocarcinoma (1) received a single 1-hour intravenous(IV) infusion of either 80 mgs or 160 mgs of the Ab1 monoclonalantibody. No further dosages of the Ab1 monoclonal antibody wereadministered to the test population.

Patients were evaluated prior to administration of the dosage, andthereafter for at least 6 weeks post dose. At the time of eachevaluation, patients were screened for plasma albumin concentration.

Results

The averaged data for both dosage concentrations (80 mgs and 160 mgs) ofthe Ab1 monoclonal antibody demonstrated an increase of about 5 g/L ofplasma albumin concentration per patient over the period of 6 weeks(FIG. 33).

What is claimed is:
 1. A method of treating a patient having a diseaseor condition associated with hypercoagulation, wherein the treatedpatient is a transplant recipient, comprises comprising administering tothe patient an interleukin-6 (IL-6) antibody or antibody fragment whichcontains the variable light chain CDRs of SEQ ID NO: 4, 5 and 6 and thevariable heavy chain CDRs of SEQ ID NO: 7, 8 or 120, and 9, whereby thepatient's coagulation profile is improved or restored to normal, andmonitoring the patient to assess coagulation profile.
 2. The method ofclaim 1, wherein said condition 9 is associated with aberrant serumlevels of one or more of D-dimer, Factor II, Factor V, Factor VIII,Factor IX, Factor XI, Factor XII, F/fibrin degradation products,thrombin-antithrombin III complex, fibrinogen, plasminogen, prothrombin,and von Willebrand factor.
 3. The method of claim 1, wherein thepatient's coagulation profile is assessed before or after treatment bymeasurement of the patient's serum level of one or more of D-dimer,Factor II, Factor V, Factor VIII, Factor IX, Factor XI, Factor XII,F/fibrin degradation products, thrombin-antithrombin III complex,fibrinogen, plasminogen, prothrombin, and von Willebrand factor.
 4. Themethod of claim 1 wherein the patient has elevated serum D-dimer levelsprior to treatment, and/or elevated C-reactive protein (CRP) levelsprior to treatment.
 5. The method of claim 1 wherein said method furthercomprises the administration of at least one statin or anti-coagulant.6. The method of claim 5 wherein said anti-IL-6 antibody or antibodyfragment further comprises a human Fc derived from IgG1, IgG2, IgG3, orIgG4.
 7. The method of claim 5 wherein the one or more anti-coagulantsare selected from abciximab, acenocoumarol, antithrombin III,argatroban, aspirin, bivalirudin, clopidogrel, dabigatran, dabigatranetexilate, desirudin, dipyridamole, eptifibatide, fondaparinux, heparin,hirudin, idraparinux, lepirudin, low molecular weight heparin,melagatran, phenindione, phenprocoumon, ticlopidine, tirofiban,warfarin, ximelagatran, ximelagatran, or any combination thereof and theone or more statins is selected from atorvastatin, cerivastatin,fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin,rosuvastatin, simvastatin, or any combination thereof.
 8. The method ofclaim 1 wherein the anti-IL-6 antibody or antibody fragment contains anFc region that has been modified to alter effector function, half-life,proteolysis, and/or glycosylation.
 9. The method of claim 1 wherein theanti-IL-6 antibody or antibody fragment is a human, humanized, singlechain or chimeric antibody.
 10. The method of claim 1 wherein theanti-IL-6 antibody or antibody fragment comprises a VH polypeptidehaving at least 90% sequence identity to SEQ ID NO: 3, 18, 19, 656, or657 and a VL polypeptide having at least 90% sequence identity to SEQ IDNO: 2, or
 20. 11. The method of claim 1 wherein the anti-IL-6 antibodyor antibody fragment is administered as part of a regimen that includesthe administration of another therapeutic compound.
 12. The method ofclaim 11 wherein the other therapeutic compound is selected from VEGFantagonists, EGFR antagonists, platins, taxols, irinotecan,5-fluorouracil, gemcitabine, leucovorine, steroids, cyclophosphamide,melphalan, vinca alkaloids, mustines, tyrosine kinase inhibitors,radiotherapy, sex hormone antagonists, selective androgen receptormodulators, selective estrogen receptor modulators, PDGF antagonists,TNF antagonists, IL-1 antagonists, interleukins, IL-12R antagonists,Toxin conjugated monoclonal antibodies, tumor antigen specificmonoclonal antibodies, Cetuximab, Bevacizumab, Pertuzumab, anti-CD20antibodies, rituximab, ocrelizumab, ofatumumab, DXL625, Trastuzumab, orany combination thereof.
 13. The method of claim 1 wherein the patient'scoagulation profile is assessed by measurement of the patient's serumlevel of D-dimer.